Molecular mechanism of seed dormancy release induced by fluridone compared with cod stratification in Notopterygium incisum.

BACKGROUND: Notopterygium incisum is an important Chinese medicinal plant. Its mature seeds have underdeveloped embryos and are physiological dormant. We found the seeds with full developed embryos can germinate after treated by fluridone (FL), an inhibitor of abscisic acid (ABA). In order to understand the molecular mechanisms underlying seed dormancy release by FL, we compared the transcriptomic changes in dormancy release induced by two different methods, FL and cold stratification (CS) in N. incisum. We further analyzed the gene expression patterns involved in seed germination and dormancy using quantitative reverse-transcription PCR. RESULTS: RNA-sequence analysis revealed more dramatic changes in the transcriptomes of FL than those in CS, particularly for genes involved in the biosynthesis and regulation of gibberellins (GAs) and ABA. The down-regulation of ABA biosynthesis genes and the dramatic up-regulation of NiCYP707As, an ABA catabolic gene, contributed to the reduced ABA levels in FL. The increased GA3 levels in CS-treated seeds were due to the up-regulation of NiGA3OX. Both NiABI5 (a positive ABA regulator) and NiGAI (a negative regulator of GA) were down-regulated in FL and CS. The upregulation of strigolactones (SLs; the metabolites with the same precursor as ABA) biosynthesis and regulatory genes in both FL- and CS-treated seeds indicates that SLs contribute positively to seed dormancy release in N. incisum. CONCLUSIONS: Our results indicated that FL- and CS-seed dormancy release possibly depends on two totally different mechanisms: alleviation of the effects of ABA and potentiation of the effects of GA, respectively. However, NiABI5 and NiGAI probably function as common factors integrating the effects of ABA and GA on seed dormancy release.

Essential oil constituents of the aerial parts and root of Cymbocarpum anethoides (Apiaceae) from Iran.

The essential oils obtained by hydrodistillation from aerial parts at different growing stages and root of Cymbocarpum anethoides DC., from Iran were investigated. The oils were analysed by GC-FID and GC-MS. Forty-five, 52, 40 and 36 components were identified in the essential oils of aerial parts (vegetative, flowering and fruit) and root representing of the 99.2, 99.0, 99.8 and 99.6% of the total oils, respectively. The essential oil of the aerial parts of the plant in vegetative stage was dominated by n-decanal (36.5%) and n-dodecanal (25.3%). n-Decanal (35.8%) and 2E-decenal (25.1%) were the main constituents of the plant oil in flowering stage whereas 2E-decenal (32.1%) and 2E-dodecenal (21.5%) were characterised as the main components of the plant oil in fruit stage. In the essential oil of root, the major identified components were 2-dodecenoic acid (29.8%) and 2E-Dodecenol (12.7%).

Cytological evaluation of Apiaceae Lindl. from Western Himalayas.

The present paper deals with cytological studies on 31 populations covering 17 species belonging to 10 genera of Apiaceae from Western Himalayas. The chromosome numbers in the two species as Chaerophyllum capnoides (n = 11) and Heracleum brunonis (n = 11), along with additional cytotypes for Pimpinella acuminata (n = 9) and Sium latijugum (n = 12) have been reported for the first time on world-wide basis. The genus Pleurospermum, although cytologically worked out earlier from outside India, its species densiflorum (n = 11) makes first representation of the genus from India. Besides, the chromosome number in Chaerophyllum aromaticum (n = 11) have been worked out for the first time from India. The course of meiosis varies from normal to abnormal in different populations of Chaerophyllum villosum, Pimpinella achilleifolia and Sium latijugum while abnormal meiotic course has been observed in all the studied populations of Chaerophyllum acuminatum, C. aromaticum, C. capnoides, Pimpinella acuminata, P. diversifolia, Pleurospermum densiflorum and Vicatia coniifolia. Such taxa are marked with meiotic abnormalities in the form of cytomixis, chromatin stickiness, formation of laggards and bridges resulting into abnormal microsporogenesis. The occurrence of structural heterozygosity has been recorded in the Chaerophyllum acuminatum and C. aromaticum. The effect of these abnormalities is clearly seen on the pollen size and fertility.

Response of Ajowan to water stress induced by polyethylene glycol (PEG) 6000 during seed germination and seedling growth.

Seed germination and subsequent metabolic changes in Ajowan (Trachyspermum ammi L.) (NRCSS AA-2) seedlings was studied under water limiting conditions, imposed by increasing concentrations of polyethylene glycol (PEG 6000). Five water stress conditions (0, -0.05, -0.1, -0.15 and -0.2 MPa) were created in the laboratory in a completely randomized design. The results revealed that water stress (-0.2 MPa) significantly reduced seed germination components like final germination percent (80%) radical (64%) and plumule (63%) length, fresh (63%) and dry (74%) weight of seedlings and vigor index (SVI) by 92% over control. Decrease in osmotic potential resulted in decreased protein content (56%) with concomitant increase in total sugars (55%) at -0.2 MPa as compared to control. Significant increase in free proline and glycine betaine content by 1.5 to 2 folds was observed at the highest water stress condition. The seedlings exhibited increased activity of superoxide dismutase and peroxidase under stressed condition. In the present study, it was found that Ajowan was a moderately drought tolerant species at laboratory level.

[Study on plant regeneration from somatic embryos of vulnerable medicinal plant Glehnia littoralis].

OBJECTIVE: To study the cause of the seeds dormancy of Glehnia littoralis in vitro and to establish plant regeneration methods via somatic embryos. METHOD: The effects of endosperm and exogenous hormone on the seed dormancy breaking of G. littoralis and the effect of hormone concentration on embryonic callus induction and plant regeneration via somatic embryos were observed, RESULTS: The germination rate of the seeds with 1/3 endosperm was the highest which achieved 31%. TDZ, 6-BA and GA3 treatment could not break seed dormancy but easily lead to abnormal seedlings. Embryogenic callus induction rates was up to 57% on MS supplemented with 1.0 mg x L(-1) 2,4-D. After 20 days culture, embryogenic calli were transferred to MS medium and cotyledonary embryos were formed in 40 days. The regenerated plants were obtained in 20 days. CONCLUSION: An effective system of plant regeneration of G. littoralis was established in this study.

Plant dispersal in the sub-Antarctic inferred from anisotropic genetic structure.

Climatic conditions and landscape features often strongly affect species' local distribution patterns, dispersal, reproduction and survival and may therefore have considerable impacts on species' fine-scale spatial genetic structure (SGS). In this study, we demonstrate the efficacy of combining fine-scale SGS analyses with isotropic and anisotropic spatial autocorrelation techniques to infer the impact of wind patterns on plant dispersal processes. We genotyped 1304 Azorella selago (Apiaceae) specimens, a wind-pollinated and wind-dispersed plant, from four populations distributed across sub-Antarctic Marion Island. SGS was variable with Sp values ranging from 0.001 to 0.014, suggesting notable variability in dispersal distance and wind velocities between sites. Nonetheless, the data supported previous hypotheses of a strong NW-SE gradient in wind strength across the island. Anisotropic autocorrelation analyses further suggested that dispersal is strongly directional, but varying between sites depending on the local prevailing winds. Despite the high frequency of gale-force winds on Marion Island, gene dispersal distance estimates (σ) were surprisingly low (<10 m), most probably because of a low pollen dispersal efficiency. An SGS approach in association with isotropic and anisotropic analyses provides a powerful means to assess the relative influence of abiotic factors on dispersal and allow inferences that would not be possible without this combined approach.

Seed longevity and germination characteristics of six fen plant species.

Fens are among the most threatened habitats in Europe as their area has decreased considerably in the last centuries. For successful management and restoration conservationists need detailed knowledge about seed bank formation and seed longevity of plants, as these features are closely related to successional and vegetation dynamical processes. I analysed seed longevity and the germination characteristics of six fen plant species by seed burial experiments. Based on seed weight, seed bank was expected for long-term persistent for the light-seeded Schoenus nigricans, Carex appropinquata, C. pseudocyperus, C. davalliana and Peucedanum palustre and also that for the medium-seeded Cicuta virosa. It was proved that, the latter two species have short-term persistent seed banks, while Carex pseudocyperus has a transient seed bank, therefore these species may only have a limited role in restoration from seed banks. It was found that Schoenus nigricans, Carex appropinquata and C. davalliana have persistent seed banks, because some of their four-year-old seeds have emerged. Fresh seeds had low germination rate in all studied species and majority of seeds emerged after winter, except for Carex pseudocyperus. After the germination peak in spring, the majority of the ungerminated seeds of Schoenus nigricans, Peucedanum palustre, Carex appropinquata, C. davalliana and Cicuta virosa entered a secondary dormancy phase that was broken in autumn. I found the seasonal emergence of the latter three species highly similar.

Deoxypodophyllotoxin, flavolignan, from Anthriscus sylvestris Hoffm. inhibits migration and MMP-9 via MAPK pathways in TNF-alpha-induced HASMC.

The matrix metalloproteinases (MMP-9 and MMP-2) in aortic smooth muscle cells (SMC) play key roles in the pathogenesis atherosclerosis. The SMC migration into the vascular wall via the bloodstream is directly linked with MMP-9 expression. Deoxypodophyllotoxin (DPT), a naturally occurring flavolignan with anti-inflammatory activity, was isolated from Anthriscus sylvestris Hoffm. and has been known inhibit the expression of MMP-9 in tumor necrosis factor-alpha (TNF-alpha) stimulated human aortic smooth muscle cells (HASMC). In this study, DPT was purified and demonstrated to inhibit the MMP-9/2 activities in TNF-alpha-induced HASMC. In addition, MMP-9 expression and migration was strongly inhibited by DPT in TNF-alpha-induced HASMC. To examine whether TNF-alpha-induced MMP-9 expressions are involved with migrations of HASMC, reverse transcription-polymerase chain reaction (RT-PCR) and luciferase-tagged promoter analysis were applied. These experiments revealed that DPT inhibited the mRNA transcription of MMP-9 gene expression. Furthermore, Western blot analysis indicated that the TNF-alpha-induced phosphorylation of extracellular signal regulated kinase 1 and 2 (ERK1/2), p38 and c-Jun N-terminal kinase (JNK) were strongly inhibited by DPT. From these results, it is concluded that DPT has an inhibitory activities on migration and MMP-2/9 activities, and MMP-9 transcription in HASMC.

[Study on rapid propagation of Changium smyrnioides].

OBJECTIVE: For the protection of Changium smyrnioides Wolff germ-plasm resources. METHODS: Rapid propagation was conducted by tissue culture. RESULTS: The best explants were leaves. The optimum culture medium for the induction of callus was MS + 2,4-D 1.0 mg/L + Kt 1.0 mg/L; that of bud was MS + 6-BA 3.0 mg/L + NAA 0.2 mg/L; and that of root was MS + IBA 0.4 mg/L or MS + NAA 0.4 mg/L. CONCLUSION: Rapid propagation of Changium smyrnioides is established with tissue culture, which provides an effective way for the sustainable utilization.

[Study on seed quality test and quality standard of Changium smyrnioides].

OBJECTIVE: To study seed quality test methods, and provide basis for establishing seed quality standard of Changium smyrnioides. METHOD: According to the seed testing regulations, the seed quality of C. smyrnioides from different producing area and in different growing years was measured. RESULT AND CONCLUSION: The range of seed purity, weight per 1000 seeds, seed moisture content, seed viability were determined and the primary seed quality classification standard was established.

[Study on seed dormancy mechanisms of Changium smyrnioides and its geminating conditions].

OBJECTIVE: To study the seed dormancy mechanisms and the optimal seed germinating conditions, so as to provide basis for the seed testing of Changium smyrnioides. METHOD: Put the seeds into wettish sand under temperature of 25, 20, 15, 10, 5 degrees C, observe the shape changing of embryo and measure its length every other 10 days. Carry on experiments of seed germination under temperature of 25,20, 15, 10, 5 degrees C on the paper bed and sand bed, determine the time of germination starting, time of germination lasting, germination rate and germination energy. RESULT: Embryo grew the most quickly under 15 degrees C. The seed germination was largely prohibited when the temperature is higher than 20 degrees C. The mildew rate was very high under 25, 20, 15, 10 degrees C. Put the seeds under 5 degrees C for 45 days could quickly break off its dormancy, and its mildew rate was lower. The seeds germinate fastest under the condition of 15-10 degrees C on the paper bed, and its germination rate was the highest. CONCLUSION: The morphological post-maturation seems the major dormancy mechanism of C. smyrnioides. Temperature is a major factor that affect seed germination of C. smymioides. The best pretreatment method is to put the seeds into 5 degrees C of wettish sand for 45 days. The best germination bed is the paper bed, and the best germination temperature is 15-10 degrees C.

Receptor-mediated increase in cytoplasmic free calcium required for activation of pathogen defense in parsley.

Transient influx of Ca(2+) constitutes an early element of signaling cascades triggering pathogen defense responses in plant cells. Treatment with the Phytophthora sojae-derived oligopeptide elicitor, Pep-13, of parsley cells stably expressing apoaequorin revealed a rapid increase in cytoplasmic free calcium ([Ca(2+)](cyt)), which peaked at approximately 1 microM and subsequently declined to sustained values of 300 nM. Activation of this biphasic [Ca(2+)](cyt) signature was achieved by elicitor concentrations sufficient to stimulate Ca(2+) influx across the plasma membrane, oxidative burst, and phytoalexin production. Sustained concentrations of [Ca(2+)](cyt) but not the rapidly induced [Ca(2+)](cyt) transient peak are required for activation of defense-associated responses. Modulation by pharmacological effectors of Ca(2+) influx across the plasma membrane or of Ca(2+) release from internal stores suggests that the elicitor-induced sustained increase of [Ca(2+)](cyt) predominantly results from the influx of extracellular Ca(2+). Identical structural features of Pep-13 were found to be essential for receptor binding, increases in [Ca(2+)](cyt), and activation of defense-associated responses. Thus, a receptor-mediated increase in [Ca(2+)](cyt) is causally involved in signaling the activation of pathogen defense in parsley.

Phytophthora parasitica elicitor-induced reactions in cells of Petroselinum crispum.

Cultured parsley (Petroselinum crispum) cells respond to treatment with elicitors derived from different species of the genus Phytophthora with transcript accumulation of defense-associated genes and the production of furanocoumarin phytoalexins. Pep-25, an oligopeptide fragment of a Phytophthora sojae 42-kDa cell wall protein, and a cell wall elicitor preparation derived from Phytophthora parasitica (Pp-elicitor) stimulate accumulation of the same gene transcripts and formation of the same pattern of furanocoumarins. Treatment of cultured cells and protoplasts with proteinase-digested Pp-elicitor identified proteinaceous constituents as active eliciting compounds in parsley. Similar to Pep- 25, Pp-elicitor induced effluxes of K+ and Cl- and influxes of protons and Ca2+. Concomitantly, as monitored in aequorin-transgenic parsley cell lines both elicitors induced an immediate increase in the cytoplasmic Ca2+ concentration up to sustained levels of 175 nM (Pp-elicitor) or 300 nM (Pep-25), respectively. The signature of the Ca2+ response differed greatly between the two elicitors tested. Extracellular Ca2+ proved essential for activation of an oxidative burst, MAP kinase activity and phytoalexin production by either elicitor. While Pp-elicitor induced a qualitatively similar spectrum of defense responses as did Pep-25, elicitor-specific quantitative differences in response intensity and kinetics suggest activation of a conserved signaling cascade through separate ligand binding sites.

Pharmacological actions of Heteromorpha trifoliata ("dombwe") on rat isolated muscle preparations.

OBJECTIVE: To investigate the effects of Heteromorpha trifoliata on rat uterine and skeletal muscle. DESIGN: Laboratory based study using experimental animals. Investigating the effects of the plant extract and agonists on isolated muscle preparations. SETTING: Department of Physiology, University of Zimbabwe. SUBJECTS: 28 Sprague-Dawley rats. MAIN OUTCOME MEASURES: Amplitude of contraction of uterine smooth muscle and skeletal muscle. RESULTS: Experiments were performed on the isolated rat uterus preparation in which strips of myometrium were placed in tissue baths filled with Kreb's solution. The aqueous extract of the root bark of Heteromorpha trifoliata ("dombwe") contracted the rat uterus. The contractions were not antagonised by atropine but were blocked by both cyproheptadine and verapamil. In addition, "dombwe" induced a contracture of the rat diaphragm muscle in the presence of alcuronium. CONCLUSIONS: The contractile effects on the uterus appear to involve stimulation of 5-HT2 receptors leading to an increase in calcium influx into the smooth muscle cell. Promotion of calcium influx could also explain the effects observed on the skeletal muscle preparation since the contracture induced by "dombwe" occurred in the presence of the nicotinic antagonist, alcuronium. In view of the effects of "dombwe" on other smooth muscle preparations (from previous work) it appears that the pharmacological profile of the crude aqueous extract of the root bark of Heteromorpha trifoliata is complex and suggestive of the presence of more than one active ingredient.