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Phytoecdysteroids in the genus Asparagus (Asparagaceae).

Phytoecdysteroids, plant steroids which are analogues of invertebrate steroid hormones, probably contribute to the deterrence of phytophagous invertebrate predators. They also seem to possess antimicrobial activity and several pharmaceutical and medicinal benefits have been ascribed to them. Here. we present a survey of seeds of 16 species of the genus Asparagus (Asparagaceae), including the crop species A. officinalis, for ecdysteroid agonists (including phytoecdysteroids) and antagonists. Seven species were found to contain ecdysteroids with levels ranging from just detectable (A. racemosus and A. sarmentosus) to relatively high (A. laricinus). RP-HPLC/RIA/bioassay has been used to separate positive extracts of four species (A. falcatus, A. laricinus, A. ramosissimus and A. scandens) and analyse the ecdysteroid profiles. The identities of the major ecdysteroids were confirmed by NP-HPLC. Seeds of A. officinalis do not contain detectable levels of ecdysteroids, but leaves, stems and roots contain low levels (detectable by RIA). This indicates that A. officinalis retains the genetic capacity to synthesise ecdysteroids and that future strategies could be developed for enhanced protection of asparagus spears through elevated ecdysteroid levels.

Anti-Infective Agents↗

Comparative genomic analyses in Asparagus.

Garden asparagus (Asparagus officinalis L.) belongs to the monocot family Asparagaceae in the order Asparagales. Onion (Allium cepa L.) and Asparagus officinalis are 2 of the most economically important plants of the core Asparagales, a well supported monophyletic group within the Asparagales. Coding regions in onion have lower GC contents than the grasses. We compared the GC content of 3374 unique expressed sequence tags (ESTs) from A. officinalis with Lycoris longituba and onion (both members of the core Asparagales), Acorus americanus (sister to all other monocots), the grasses, and Arabidopsis. Although ESTs in A. officinalis and Acorus had a higher average GC content than Arabidopsis, Lycoris, and onion, all were clearly lower than the grasses. The Asparagaceae have the smallest nuclear genomes among all plants in the core Asparagales, which typically have huge genomes. Within the Asparagaceae, European Asparagus species have approximately twice the nuclear DNA of that of southern African Asparagus species. We cloned and sequenced 20 genomic amplicons from European A. officinalis and the southern African species Asparagus plumosus and observed no clear evidence for a recent genome doubling in A. officinalis relative to A. plumosus. These results indicate that members of the genus Asparagus with smaller genomes may be useful genomic models for plants in the core Asparagales.

Asparagus Plant↗

Two pyrrolo[1,2-a]azepine type alkaloids from Stemona collinsae Craib: structure elucidations, relationship to asparagamine A, and a new biogenetic concept of their formation.

The alkaloids 1',2'-didehydrostemofoline (2) and 2'-hydroxystemofoline (3) from Stemona collinsae Craib (Stemonaceae) were studied by X-ray crystallography and NMR spectroscopy, and they are compared with the parent compound stemofoline (1). The X-ray analysis of the CH2Cl2 solvate of 2'-hydroxystemofoline (3) allowed the determination of the absolute configuration of this compound unequivocally, whereas optical rotation was used to infer the absolute configuration of 1',2'-didehydrostemofoline (2). Based on these results, it is shown that asparagamine A isolated from Asparagus racemosus Willd. (Asparagaceae) is identical to 1',2'-didehydrostemofoline obtained from S. collinsae Craib, and that the reported plant source of asparagamine A was most likely a Stemona species. In the context of the current investigations, a novel concept on the biosynthesis of Stemona alkaloids has been worked out and is presented here.

Alkaloids↗

The rice genome encodes two vacuolar invertases with fructan exohydrolase activity but lacks the related fructan biosynthesis genes of the Pooideae.

* Fructans are believed to contribute to cold and drought tolerance in several plant families (Poaceae, Asparagaceae and Asteraceae), but it is not clear why the ability to accumulate these polymers is found in some genera (e.g. Triticum) but not in others (e.g. Oryza). * As fructan biosynthesis enzymes (FBEs) evolved from vacuolar invertases (VINs), we searched the rice genome sequence for genes related to both FBE and VIN genes of wheat and other members of the Pooideae. We compared them at the levels of exon-intron structure, protein sequence, and the enzymatic properties of recombinant proteins after expression in the yeast Pichia pastoris. * We found that rice possesses two VIN genes (OsVIN1 and OsVIN2) and no FBE genes. FBE genes appear to have arisen in the Pooideae by a series of gene duplications from an ancestor of wheat TaVIN3. Recombinant TaVIN2, OsVIN1 and OsVIN2 behaved as invertases with no FBE activity, but possessed high fructan exohydrolase activity, especially OsVIN1. * The engineering of fructan accumulation into rice for greater stress tolerance could founder on endogenous exohydrolases, but the fact that OsVIN1 transcripts are absent from peduncles of well watered and drought-stressed plants removes one potential obstacle to this endeavour.

Amino Acid Sequence↗

Methods for characterizing plant fibers.

The effectiveness of different microscopy techniques for measuring the dimensions of ultimate fibers from harakeke (Phormium tenax, New Zealand flax) was investigated using a factorial experimental design. Constant variables were geographical location, location of specimens along the leaf, season (winter), individual plant, a fourth leaf from a north-facing fan, age of plant, and cultivars (two). Experimental variables were microscopy techniques and measurement axis. Measurements of width and length of harakeke ultimate fibers depended on the microscopic preparation/technique used as well as the cultivar examined. The best methods were (i) transverse sections of leaf specimens 4 microm thick, embedded in Paraplast and observed using light microscopy, and (ii) nonfixed ultimate fibers observed using scanning electron microscopy.

Asparagaceae↗

Antifungal activity of tuberose absolute and some of its constituents.

The antifungal activity of the absolute of tuberose (Polianthes tuberosa ) and some of its constituents were evaluated against the mycelial growth of Colletotrichum gloeosporioides on potato-dextrose-agar medium. Tuberose absolute showed only mild activity at a concentration of 500 mg/L. However, three constituents present in the absolute, namely geraniol, indole and methyl anthranilate exhibited significant activity showing total inhibition of the mycelial growth at this concentration.

Antifungal Agents↗

Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceum Baker--an endangered medicinal herb.

An efficient in vitro multiplication system via multiple shoot bud induction and regeneration has been developed in Chlorophytum arundinaceum using shoot crown explants. Optimum regeneration frequency (87%) and desirable organogenetic response in the form of de novo organized multiple shoot buds without an intervening callus phase was obtained on Murashige and Skoog's (MS) minimal organics medium containing 3% sucrose (w/v) supplemented with 4 x 10(-6) M Kn and 2 x 10(-6) MIBA. Axenic secondary explants with multiple shoot buds on subculturing elicited best response with 1 x 10(-5) M Kinetin (Kn) and 5 x 10(-6) M indole-3-butyric acid (IBA) giving rise to an average of 18.74 shoots per culture with mean shoot length of 7.6 cm +/- 1.73. Varying molar ratios of either Kn/IBA or Kn/NAA revealed statistically significant differences in the regeneration frequencies among the phytohormone treatments. It was observed that the shoot bud differentiation and regeneration was influenced by the molar ratios of cytokinins/auxin rather than their relative concentrations. Healthy regenerated shoots were rooted in half strength MS basal medium containing 3% sucrose (w/v) supplemented with 5 x 10(-6) M IBA. Following simple hardening procedures, rooted plantlets, were transferred to soil-sand (1:1; v/v) with more than 90% success. Genetic fidelity was assessed using random amplified polymorphic DNA (RAPD), karyotype analysis and meiotic behaviour of in vitro and in vivo plants. Five arbitrary decamers displayed same banding profile within all the micropropagated plants and in vivo explant donor. The cytological and molecular analysis complemented and compared well and showed no genomic alterations in the plants regenerated through shoot bud differentiation. High multiplication frequency, molecular, cytological and phenotypic stability ensures the efficacy of the protocol developed for the production and conservation of this important endangered medicinal herb.

Asparagaceae↗

Effects of an increase in summer precipitation on leaf, soil, and ecosystem fluxes of CO2 and H2O in a sotol grassland in Big Bend National Park, Texas.

Global climate models predict that in the next century precipitation in desert regions of the USA will increase, which is anticipated to affect biosphere/atmosphere exchanges of both CO(2) and H(2)O. In a sotol grassland ecosystem in the Chihuahuan Desert at Big Bend National Park, we measured the response of leaf-level fluxes of CO(2) and H(2)O 1 day before and up to 7 days after three supplemental precipitation pulses in the summer (June, July, and August 2004). In addition, the responses of leaf, soil, and ecosystem fluxes of CO(2) and H(2)O to these precipitation pulses were also evaluated in September, 1 month after the final seasonal supplemental watering event. We found that plant carbon fixation responded positively to supplemental precipitation throughout the summer. Both shrubs and grasses in watered plots had increased rates of photosynthesis following pulses in June and July. In September, only grasses in watered plots had higher rates of photosynthesis than plants in the control plots. Soil respiration decreased in supplementally watered plots at the end of the summer. Due to these increased rates of photosynthesis in grasses and decreased rates of daytime soil respiration, watered ecosystems were a sink for carbon in September, assimilating on average 31 mmol CO(2) m(-2) s(-1) ground area day(-1). As a result of a 25% increase in summer precipitation, watered plots fixed eightfold more CO(2) during a 24-h period than control plots. In June and July, there were greater rates of transpiration for both grasses and shrubs in the watered plots. In September, similar rates of transpiration and soil water evaporation led to no observed treatment differences in ecosystem evapotranspiration, even though grasses transpired significantly more than shrubs. In summary, greater amounts of summer precipitation may lead to short-term increased carbon uptake by this sotol grassland ecosystem.

Asparagaceae↗

A new potyvirus from tuberose ( Polianthes tuberosa) in China.

Tuberose plants with mild mottle symptoms, growing in a glasshouse in Hangzhou, China, contained virions and inclusion bodies typical of a potyvirus. The virus was mechanically transmitted to tuberose but not to 14 other test plant species. A fragment of 4607 nucleotides, corresponding to the 3'-half of a typical potyvirus was amplified by RT-PCR using degenerate primers and sequenced. The most similar sequence in the databases was that of Tuberose mild mosaic virus (TuMMV) from Taiwan and this was the only virus significantly related to it in phylogenetic analyses. The new sequence had 71.1% nt and 76.6% aa identity to TuMMV in the coat protein. Western blot analyses using antisera raised to expressed coat protein showed that the two viruses were serologically related. Although there are no substantial biological data to distinguish the Hangzhou isolate from TuMMV, the molecular difference between the two virus isolates is similar to, or slightly greater than, that between several pairs of well-established potyvirus species. These results therefore suggest that the Hangzhou isolate should be regarded as a new member of the genus Potyvirus, and we have tentatively named it Tuberose mild mottle virus.

Amino Acid Sequence↗

Variations in the fibre repeat between samples of cellulose I from different sources.

A powder X-ray diffractometer was used to measure the fibre repeat in cellulose I with sufficient precision to detect variations between samples from different sources. The variations were correlated with the lateral dimensions of the crystallites and were attributed to different minimum-energy fibre repeats for chains in the interiors and on the surfaces of crystallites. Results were interpreted in terms of a model for internal mechanical stress in which the interior chains were under compression and the surface chains under tension to ensure identical fibre repeats for all chains. The model was used to extrapolate the fibre repeat to a value of 1.043 nm for a hypothetical, infinitely large crystal, and to 1.029 nm for a crystallite so narrow that all chains were exposed on surfaces.

Animals↗

Saponins from Furcraea selloa var. marginata.

Four steroidal saponins were isolated from the leaves of Furcraea selloa var. marginata. These included one furostanol saponin, furcreafurostatin (1), and three known spirostanol saponins, furcreastatin (3), yuccaloeside C (4) and cantalasaponin-1 (5). The 22-O-methyl ether (2) of furcreafurostatin (1) was also characterized. The structures were determined by using a combination of spectroscopic techniques.

Asparagaceae↗

Free radical scavenging potential of Chlorophytum tuberosum Baker.

Chlorophytum tuberosum Baker commonly referred as 'Musli' has been widely used as a potent 'Rasayana' drug in 'Ayurveda' as a rejuvenator and tonic. Antioxidant potential of Chlorophytum tuberosum has been investigated for their ability to scavenge 1,1,diphenyl picryl hydrazyl (DPPH), nitric oxide radical along with their capacity to reduce lipid peroxidation in rat liver homogenate, chelation of ferrous ion, radical scavenging potential using chemiluminescence and their total antioxidant capacity. Sugar, starch, protein, and Vitamin C content were estimated spectrophotometrically along with the percentages of the individual amino acids by HPLC and individual sugars by using HPTLC as standardization tool. The extract has been found to possess antioxidant activity in all the models tested as evident by IC50 values being 225.31, 888.44, 809.22 and 422.97 microg/ml for scavenging of DPPH, nitric oxide, lipid peroxidation and ferry bi-pyridyl complex, respectively, along with a integral anitoxidant activity of 2.986 nmol ascorbic acid/g equivalents in photochemiluminescence assay.

Animals↗

Species of colletotrichum on agavaceae.

Species of Colletotrichum cause diseases on a wide range of hosts, frequently infecting plants in the Agavaceae (monocotyledons: Liliales). Three species of Colletotrichum restricted to the Agavaceae were detected through morphological studies of specimens and molecular sequence analyses of the LSU of the nu-rDNA and the ITS region of the nu-rDNA from cultures. Colletotrichum agaves on Agave is fully described and illustrated. Colletotrichum dracaenophilum is described as a new species for isolates having long conidia and occurring on Dracaena sanderiana from China. Colletotrichum phormii and Glomerella phormii are determined to be the correct scientific names for the asexual and sexual states, respectively, of a species commonly referred to as C. rhodocyclum and G. phacidiomorpha occurring mainly on Phormium. In addition, C. gloeosporioides and C. boninense were isolated from plants in the Agavaceae. All species of Colletotrichum described on Agavaceae were evaluated based on type specimens. A key to the five species of Colletotrichum on Agavaceae is included. This paper includes one new species, Colletotrichum dracaenophilum, and three new combinations, Colletotrichum phormii, Glomerella phormii, and Phaeosphaeriopsis phacidiomorpha.

Asparagaceae↗

Phytochemistry and antimycobacterial activity of Chlorophytum inornatum.

In a project to investigate plant derived natural products from the Liliaceae with activity against fast-growing strains of mycobacteria, we have identified two new metabolites from Chlorophytum inornatum. The active principle, a new homoisoflavanone (1) was identified as 3-(4'-methoxybenzyl)-7,8-methylenedioxy-chroman-4-one. The metabolite assigned as 7-(1'-hydroxyethyl)-2-(2''-hydroxyethyl)-3,4-dihydrobenzopyran (2) was characterised by extensive 1- and 2D NMR spectroscopy. The antimycobacterial activity of this plant was mainly due to the homoisoflavonoid which exhibited minimum inhibitory values ranging from 16-256 microg/ml against four strains of fast-growing mycobacteria.

Anti-Bacterial Agents↗

Comparative analysis of the plasmids from two isolates of "Candidatus Phytoplasma australiense".

Two plasmids from the plant-pathogenic mollicute "Candidatus Phytoplasma australiense" were completely sequenced from two isolates derived from different plant hosts. Plasmid pPAPh2 (3607bp) was obtained from Phormium showing Phormium yellow leaf symptoms and pPASb11 (3635bp) from strawberry showing strawberry lethal yellows symptoms. The plasmids varied in their copy number and nucleotide sequence yet contained the same four open reading frames (ORFs). The deduced amino acid sequence derived from ORF1 shares similarity with hypothetical proteins encoded on the plasmids from onion yellows and beet leafhopper-transmitted virescence agent phytoplasmas. The deduced amino acid sequences of both ORF2 and ORF3 share similarity with functionally unknown proteins on the chromosome of onion yellows phytoplasma. An ORF with a similar sequence to ORF2 is also present on the chromosome of "Ca. P. australiense." The deduced amino acid sequence derived from ORF4 is most similar to replication proteins encoded by other phytoplasma plasmids and by geminiviruses, the only protein on the plasmids for which a putative function can be assigned. The identities of the deduced amino acid sequences of ORF1, ORF2, ORF3, and ORF4 between pPAPh2 and pPASb11 were 89, 68, 91, and 68%, respectively; the differences being consistent with the subgroup status of the parental phytoplasmas.

Amino Acid Sequence↗

Spirostanol pentaglycosides from the underground parts of polianthestuberosa.

Phytochemical analysis of the underground parts of Polianthes tuberosa has resulted in the isolation of four new spirostanol saponins with five monosaccharides (1-4). Their structures were determined by spectroscopic analysis, including extensive 1D and 2D NMR data, and the results of hydrolytic cleavage. The cytotoxic activities of 1-4 against HL-60 human promyelocytic leukemia cells and HSC-2 human oral squamous cell carcinoma cells are reported.

Antineoplastic Agents, Phytogenic↗

Spirostanol and furostanol glycosides from the fresh tubers of Polianthes tuberosa.

Six new steroid glycosides--two spirostanols, polianthosides B and C (1, 2), and four furostanols, polianthosides D-G (3-6)--were isolated from the fresh tubers of Polianthes tuberosa, together with seven known spirostanols (7-13) and a known furostanol (14) saponins. Their structures were elucidated on the basis of spectroscopic analysis and the results of acidic and enzymatic hydrolysis. The cytotoxic activities of 1-14 against HeLa cells are reported.

Antineoplastic Agents, Phytogenic↗

Action of (2-->1)Fructo-oligopolysaccharide fraction of Chlorophytum borivilianum against Streptozotocin-Induced oxidative stress.

A fructo-oligosaccharide was isolated from Chlorophytum borivilianum and identified as O-beta-D-fructofuranosyl-(2-->1)-(beta-D-fructofuranosyl) (n)-(2-->1)-alpha-D-glucopyranoside (n = 5 - 30) using high-pressure anion exchange chromatography, MALDI-MS, NMR, GC, HPTLC and chemical analysis. The extract and the fructo-oligosaccharide were found to have significant antidiabetic activity with the blood sugar levels being 118.32 +/- 3.56 and 110.21 +/- 4.22, respectively, as compared to the control value of 231.25 +/- 3.03 along with moderate antioxidant activity in streptozotocin-induced diabetic animals.

Animals↗