Traceability the provenience of cultivated Paris polyphylla Smith var. yunnanensis using ATR-FTIR spectroscopy combined with chemometrics.

The conventional procedures, based on attenuated total reflectance-Fourier transform infrared spectrometry (ATR-FTIR), have been developed for the origins traceability of cultivated Paris polyphylla Smith var. yunnanensis (PPY) samples with the help of partial least square discriminant analysis (PLS-DA) and random forest. In this study, a set of 219 batch cultivated PPY samples, containing the cultivation years of 5, 6 and 7, and covering the municipal districts of Chuxiong, Dali, Honghe, Lijiang and Yuxi in Yunnan Province, China, were used to build the discrimination models. Firstly, a visualized analysis was carried out by t-distributed stochastic neighbor embedding (t-SNE) to reduce each data point in a two-dimensional map and make a knowledge of the sample distribution tendency. Secondly, the single spectra data sets of Paridis rhizome and leaf tissues, and the combination of these two data sets with variable selection (mid-level data fusion strategy), were used to establish PLS-DA and random forest models, and parallelly compared the model performance. Results demonstrated that the discrimination ability of PLS-DA preceded the random forest model, and the classification performance was remarkably improved after mid-level data fusion. These results verified each other by 5-, 6- and 7-year old Paridis samples and indicated that the model performance established in the present study was reliable. Besides, five agronomic characters, including the plant height, dry weight of rhizome and leaf tissues, and the allocation of rhizome and leaf were determined and analyzed, results of which indicated that the dry weight and their allocation was significantly different among various origins and fluctuated with the cultivation years. This study was using a comprehensive and green analytical method to discriminate the cultivated Paridis according to their provenances, which was simultaneously benefited for the appropriate cultivation areas selection based on the dry weight of rhizome tissues.

[Exploring the clinical characters of Shugan Jieyu capsule through text mining].

In this paper,the potential climate factors affecting the Pairs polyphylla var. yunnanensis distribution in China at rational scales were selected from related literatures, using the sampling point geographic information from of P. polyphylla var. yunnanensis, combine the maximum entropy model (MaxEnt) with spatial analyst function of ArcGIS software, to study the climate suitability of P. polyphylla var. yunnanensis cultivating region in China and the leading climate factors. The results showed that, average rainfall in August, average rainfall in October, coefficient of variation of seasonal precipitation, the average temperature of the dry season, isothermal characteristic, average temperature in July were the leading climate factors affecting the potential distribution of P. polyphylla var. yunnanensis cultivating region in China, with their cumulative contribution rate reached 97.2% of all candidate climate factors. Existence probability of the region to be predicted of P. polyphylla var. yunnanensis through the constructed model, the climate unsuitable region, low, medium and high region of P. polyphylla var. yunnanensis in China were clarified and the threshold of climatic factors were gave and clarified the climate characteristics of the cultivating region in each climatic suitability division. The results of research can provide reference for production layout and introduction of P. polyphylla var. yunnanensis.

Analysis of the floral MADS-box genes from monocotyledonous Trilliaceae species indicates the involvement of SEPALLATA3-like genes in sepal-petal differentiation.

The evolution of greenish sepals from petaloid outer tepals has occurred repeatedly in various lineages of non-grass monocots. Studies in distinct monocot species showed that the evolution of sepals could be explained by the ABC model; for example, the defect of B-class function in the outermost whorl was linked to the evolution of sepals. Here, floral MADS-box genes from three sepal-bearing monocotyledonous Trilliaceae species, Trillium camschatcense, Paris verticillata, and Kinugasa japonica were examined. Unexpectedly, expression of not only A- but also B-class genes was detected in the sepals of all three species. Although the E-class gene is generally expressed across all floral whorls, no expression was detected in sepals in the three species examined here. Overexpression of the E-class SEPALLATA3-like gene from T. camschatcense (TcamSEP) in Arabidopsis thaliana produced phenotypes identical to those reported for orthologs in other monocots. Additionally, yeast hybrid experiments indicated that TcamSEP could form a higher-order complex with an endogenous heterodimer of B-class APETALA3/DEFICIENS-like (TcamDEF) and PISTILLATA/GLOBOSA-like (TcamGLO) proteins. These results suggest a conserved role for Trilliaceae SEPALLATA3-like genes in functionalization of the B-class genes, and that a lack of SEPALLATA3-like gene expression in the outermost whorl may be related to the formation of greenish sepals.

Responses of two understory herbs, Maianthemum canadense and Eurybia macrophylla, to experimental forest warming: early emergence is the key to enhanced reproductive output.

PREMISE OF THE STUDY: Understory herbs might be the most sensitive plant form to global warming in deciduous forests, yet they have been little studied in the context of climate change. METHODS: A field experiment set up in Minnesota, United States simulated global warming in a forest setting and provided the opportunity to study the responses of Maianthemum canadense and Eurybia macrophylla in their natural environment in interaction with other components of the ecosystem. Effects of +1.7° and +3.4°C treatments on growth, reproduction, phenology, and gas exchange were evaluated along with treatment effects on light, water, and nutrient availability, potential drivers of herb responses. KEY RESULTS: Overall, growth and gas exchanges of these two species were modestly affected by warming. They emerged up to 16 (E. macrophylla) to 17 d (M. canadense) earlier in the heated plots than in control plots, supporting early-season carbon gain under high light conditions before canopy closure. This additional carbon gain in spring likely supported reproduction. Eurybia macrophylla only flowered in the heated plots, and both species had some aspect of reproduction that was highest in the +1.7°C treatment. The reduced reproductive effort in the +3.4°C plots was likely due to reduced soil water availability, counteracting positive effects of warming. CONCLUSIONS: Global warming might improve fitness of herbaceous species in deciduous forests, mainly by advancing their spring emergence. However, other impacts of global warming such as drier soils in the summer might partly reduce the carbon gain associated with early emergence.

Effect of plastic mulching on mycotoxin occurrence and mycobiome abundance in soil samples from asparagus crops.

Plastic mulching (PM) is widely used in modern agriculture because of its advantageous effects on soil temperature and water conservation, factors which strongly influence the microbiology of the soil. The aim of this study was to assess the effect of PM on mycotoxin occurrence in relation with mycobiome abundance/diversity and soil physicochemical properties. Soil samples were collected from green (GA) and white asparagus (WA) crops, the last under PM. Both crops were cultivated in a ridge-furrow-ridge system without irrigation. Samples were analyzed for mycotoxin occurrence via liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS). Total colony-forming unit was indicative of mycobiome abundance, and analysis of mycobiome diversity was performed by internal transcribed spacer (ITS) sequencing. PM avoided the drop of soil temperature in winter and allowed higher soil temperature in early spring compared to non-covered soil. Moreover, the use of PM provided controlled conditions for water content in soil. This was enough to generate a dissimilar mycotoxin occurrence and mycobiome diversity/abundance in covered and non-covered soil. Mycotoxin soil contamination was confirmed for deoxynivalenol (DON), range LOD to 32.1 ng/g (LOD = 1.1 ng/g). The DON values were higher under PM (average 16.9 ± 10.1 ng/g) than in non-covered soil (9.1 ± 7.9 ng/g); however, this difference was not statically significant (p = 0.09). Mycobiome analysis showed a fungal compartment up to fivefold higher in soil under PM compared to GA. The diversity of the mycobiome varied between crops and also along the soil column, with an important dominance of Fusarium species at the root zone in covered soils.

Dissecting the role of two cytokinin analogues (INCYDE and PI-55) on in vitro organogenesis, phytohormone accumulation, phytochemical content and antioxidant activity.

There is a continuous search for new chemical entities to expand the collection of suitable compounds to increase the efficiency of micropropagation protocols. Two cytokinin (CK) analogues, 2-chloro-6-(3-methoxyphenyl)aminopurine (INCYDE) and CK antagonist 6-(2-hydroxy-3-methylbenzylamino)purine (PI-55) were used as a tool to elucidate the auxin-CK crosstalk under in vitro conditions in the medicinally important plant, Eucomis autumnalis subspecies autumnalis. These compounds were tested at 0.01, 0.1 and 10 µM alone as well as in combination with benzyladenine (BA) and naphthaleneacetic acid (NAA). The organogenesis, phytohormone content, phytochemical and antioxidant response in 10 week-old-in vitro regenerated E. autumnalis subspecies autumnalis was evaluated. INCYDE generally favoured shoot regeneration while the effect of PI-55 was more evident in root proliferation. Overall, INCYDE promoted the accumulation of higher concentrations and varieties of endogenous CK relative to the PI-55 treatments. In contrast, higher concentration of indole-3-acetic acid and 2-oxindole-3-acetic acid were generally observed in PI-55-supplemented cultures when compared to plantlets derived from INCYDE. Both CK analogues (individually and in-conjunction with exogenously applied PGRs) significantly influenced the phytochemicals and consequently the antioxidant potential of the in vitro regenerants. These results provided insight on how to alleviate root inhibition, a problem which causes considerable loss of several elite species during micropropagation.

Physiological role of phenolic biostimulants isolated from brown seaweed Ecklonia maxima on plant growth and development.

MAIN CONCLUSION: Eckol, a major phenolic compound isolated from brown seaweed significantly enhanced the bulb size and bioactive compounds in greenhouse-grown Eucomis autumnalis. We investigated the effect of eckol and phloroglucinol (PG) (phenolic compounds) isolated from the brown seaweed, Ecklonia maxima (Osbeck) Papenfuss on the growth, phytochemical and auxin content in Eucomis autumnalis (Mill.) Chitt. The model plant is a popular medicinal species with increasing conservation concern. Eckol and PG were tested at 10(-5), 10(-6) and 10(-7) M using soil drench applications. After 4 months, growth parameters, phytochemical and auxin content were recorded. When compared to the control, eckol (10(-6) M) significantly improved bulb size, fresh weight and root production while the application of PG (10(-6) M) significantly increased the bulb numbers. However, both compounds had no significant stimulatory effect on aerial organs. Bioactive phytochemicals such as p-hydroxybenzoic and ferulic acids were significantly increased in eckol (10(-5) M) and PG (10(-6) M) treatments, compared to the control. Aerial (1,357 pmol/g DW) and underground (1,474 pmol/g DW) parts of eckol-treated (10(-5) M) plants yielded the highest concentration of indole-3-acetic acid. Overall, eckol and PG elicited a significant influence on the growth and physiological response in E. autumnalis. Considering the medicinal importance of E. autumnalis and the increasing strains on its wild populations, these compounds are potential tools to enhance their cultivation and growth.

Karyological relationships in Indian species of Drimia based on fluorescent chromosome banding and nuclear DNA amount.

The genus Drimia (syn. Urginea), commonly called squill, represents a species complex, infrageneric delimitation being ill-defined due to morphological variability, population variation within species and polyploidy. In the present study, fluorescent chromosome banding and measurements of nuclear DNA content by flow cytometry were performed in five Indian species of Drimia: Drimia indica, Drimia polyantha, Drimia razii, Drimia wightii and Drimia coromandeliana to elucidate taxonomic relationship and obtain possible insights into the evolutionary processes within this group. All taxa analyzed exhibited similar karyomorphology with subtle differences accounted by nucleolar chromosomes. Nuclear DNA content ranged from 20.41 pg/2C in D. polyantha to 40.80 pg/2C in D. coromandeliana and was positively correlated with chromosome number (r = 0.67, P = 0.02) and total diploid chromatin length (r = 0.59, P = 0.06). Fluorescent chromosome banding revealed the presence of CMA(+ve)/DAPI(-ve) signals associated with nucleolar chromosomes presumably coincident with NOR in all species and unique CMA(+ve) signals in diploid populations of D. indica. Satellite polymorphism between homologous NOR-bearing chromosomes was observed which supports hybrid origin of the taxon. UPGMA dendrogram and scatter diagrams based on karyological parameters indicated a close relationship of D. indica, D. razii and D. polyantha while D. wightii and D. coromandeliana appeared distant. D. wightii appeared more close to D. indica than to all other species based on genome size and karyomorphology. As a whole, D. indica showed high intra-specific variability with populations exhibiting intergrading characters with other species. In conclusion, it is likely that hybridization followed by reproductive isolation of polymorphic forms arising by adaptation to different ecological niches resulted in species diversification of Drimia in India, probably from a common ancestor similar to D. indica.

[Phenotypic Trait Variation, Principal Component, Correlation and Path Analysis of Paris polyphylla var. yunnanensis].

OBJECTIVE: To investigate the phenotypic trait variation range of Paris polyphylla var. yunnanensis, and to look for phenotypic traits closely related with yield, in order to provide reference for its breeding in the future. METHODS: Wild plants of Paris polyphylla var. yunnanensis populations cultivated for three years in Xishuangbanna, Yunnan Province were chosen. The plants of Paris polyphylla var. yunnanensis were randomly labelled, its phenotypic traits were observed and analyzed by variation, principal component, correlation, variance and path analysis. RESULTS: Eleven phenotypic traits variation of the populations of Paris polyphylla var. yunnanensis were great, mainly distributed in yield, growth and genetic characteristics. Phenotypic trait of different growth years (4 - 10 years) including stem height, stem diameter, flower stem height, flower stem diameter, leaf number, leaf length and leaf width had no significant difference(P > 0. 05). The underground phenotypic traits including root weight, root length and root diameter increased significant (P <0. 05) with plant growth years. Effect of the other phenotypic traits on root weight was as follows: root diameter (0. 2550) > root growth years (0. 1183 ) > root length(0. 0202) > stem diameter(0. 0081) > stem height (0. 0044) > leaf number (0. 0025) > leaf length (0. 0018) > leaf width (0. 0004) > flower stem height (0. 0003) > flower stem diameter (0. 0001). CONCLUSION: The phenotypic traits on yield, growth and genetic characteristics have rich variations, and the phenotypic traits on the ground have no relationship with plant growth years, the plant growth years mainly affect the underground phenotypic traits. When choosing different root types, root diameter can be used as the preferred character, stem diameter as the second preferred traits when the root traits information not be easily obtained.

[Research progress on enlargement of medicinal resources of Paridis Rhizome].

Currently, as an important raw material of Chinese traditional patent medicines, Paridis Rhizome is in great demand, which led to its price increases. In order to protect the wild resources and satisfy market demand of Paridis rhizome, the researches in various directions were conducted, involved its chemical composition, pharmacological action, clinical application, resource investigation, artificial cultivation, etc. Herein, the chemical studies of genus Paridis Rhizome, aerial parts of Paridis Rhizome gummy and starchy Paridis Rhizome, and the studies of endophyte in Paridis Rhizome were reviewed and analyzed in order to explore the substitutes of Paridis Rhizome, and provide the reference for the enlargement of medicinal resources of Paridis Rhizome. It manifests that the steroidal saponins, the important chemical compositions in Paridis Rhizome were tested in genus Paridis Rhizome, aerial parts of Paridis Rhizome, gummy Paridis Rhizome and the endophyte in Paridis Rhizome. However, the further experimental studies and clinical verification works should be carried out to confirm the final substitute.

[Effects of mycorrhizal colonization and medicine quality of Paris polyphylla var. yunnanensis inoculated by different foreign AM fungi species].

After 28 foreign species of AM fungi were inoculated in sterilized soil, the effects of the AM mycorrhizal colonization and the medicine quality of Paris polyphylla var. yunnanensis were observed by combination of inoculation test in pot at room temperature and instrumental analysis. The results showed that, compared with control group (CK), the inoculation of foreign AM fungi in the soil influenced the spore density, mycorrhizal infection rate, and colonization intensity of AM fungi in root system of P. polyphylla var. yunnanensis. The inoculation of foreign AM fungi enhanced the mycorrhiza viability of P. polyphylla var. yunnanensis by increasing the activity of succinic dehydrogenase (SDH) and alkaline phosphatase (ALP) in intraradical hyphae. The content of single steroid saponin in rhizome of P. polyphylla var. yunnanensis showed variation after P. polyphylla var. yunnanensis was inoculated by different foreign species of AM fungi, which was beneficial for increasing the medicine quality; however, the kinds of steroid saponin showed no difference. In a degree, there was a selectivity of symbiosis between P. polyphylla var. yunnanensis and foreign AM fungi. And we found that the Claroideoglomus claroideum and Racocetra coralloidea were best foreign AM fungi species for cultivating P. polyphylla var. yunnanensis under field condition.

Improved growth and colchicine concentration in Gloriosa superba on mycorrhizal inoculation supplemented with phosphorus-fertilizer.

BACKGROUND: Gloriosa superba produces an array of alkaloids including colchicine, a compound of interest in the treatment of various diseases. The tuber of Gloriosa superba is a rich source of colchicine which has shown anti-gout, anti-inflammatory, and anti-tumor activity. However, this promising compound remains expensive and Gloriosa superba is such a good source in global scale. Increase in yield of naturally occurring colchicine is an important area of investigation. MATERIALS AND METHODS: The effects of inoculation by four arbuscular mycorrhizal (AM), fungi, Glomus mossae, Glomus fasciculatum, Gigaspora margarita and Gigaspora gilmorei either alone or supplemented with P-fertilizer, on colchicine concentration in Gloriosa superba were studied. The concentration of colchicine was determined by high-performance thin layer chromatography. RESULTS: The four fungi significantly increased concentration of colchicine in the herb. Although there was significant increase in concentration of colchicine in non-mycorrhizal P-fertilized plants as compared to control, the extent of the increase was less compared to mycorrhizal plants grown with or without P-fertilization. This suggests that the increase in colchicine concentration may not be entirely attributed to enhanced P-nutrition and improved growth. Among the four AM fungi Glomus mossae was found to be best. The total colchicine content of plant (mg / plant) was significantly high in plants inoculated with Glomus mossae and 25 mg kg(-1)phosphorus fertilizer (348.9 mg /plant) while the control contain least colchicine (177.87 mg / plant). CONCLUSION: The study suggests a potential role of AM fungi in improving the concentration of colchicine in Gloriosa superba tuber.

Intermediate complex morphophysiological dormancy in seeds of the cold desert sand dune geophyte Eremurus anisopterus (Xanthorrhoeaceae; Liliaceae s.l.).

BACKGROUND AND AIMS: Little is known about morphological (MD) or morphophysiological (MPD) dormancy in cold desert species and in particular those in Liliaceae sensu lato, an important floristic element in the cold deserts of Central Asia with underdeveloped embyos. The primary aim of this study was to determine if seeds of the cold desert liliaceous perennial ephemeral Eremurus anisopterus has MD or MPD, and, if it is MPD, then at what level. METHODS: Embryo growth and germination was monitored in seeds subjected to natural and simulated natural temperature regimes and the effects of after-ripening and GA3 on dormancy break were tested. In addition, the temperature requirements for embryo growth and dormancy break were investigated. KEY RESULTS: At the time of seed dispersal in summer, the embryo length:seed length (E:S) ratio was 0·73, but it increased to 0·87 before germination. Fresh seeds did not germinate during 1 month of incubation in either light or darkness over a range of temperatures. Thus, seeds have MPD, and, after >12 weeks incubation at 5/2 °C, both embryo growth and germination occurred, showing that they have a complex level of MPD. Since both after-ripening and GA3 increase the germination percentage, seeds have intermediate complex MPD. CONCLUSIONS: Embryos in after-ripened seeds of E. anisopterus can grow at low temperatures in late autumn, but if the soil is dry in autumn then growth is delayed until snowmelt wets the soil in early spring. The ecological advantage of embryo growth phenology is that seeds can germinate at a time (spring) when sand moisture conditions in the desert are suitable for seedling establishment.

Simultaneous determination of bufadienolides and phenolic compounds in sea squill (Drimia maritima (L.) Stearn) by HPLC-DAD-MSn as a means to differentiate individual plant parts and developmental stages.

Mediterranean sea squill (Drimia maritima (L.) Stearn) is used in the production of medicinal products. Current HPLC methods comprise tedious sample clean-up and have been merely focused on the analysis of cardiac glycosides, whereas a thorough characterization of D. maritima considering both the latter compound class and more hydrophilic secondary metabolites in one HPLC run has not been performed so far. Consequently, a novel HPLC-DAD-MS(n) method has been developed allowing the simultaneous determination of both cardiac glycosides and phenolic compounds, which is characterized by simplified sample preparation. This method was applied to characterize sea squill, revealing a complex profile of its extractive compounds derived from the two classes. Furthermore, the potential of the method reported here to quantitate the predominant compounds, i.e., dihydroquercetin derivatives and bufadienolides, was demonstrated. The occurrence of phenolic compounds, not described for sea squill so far, and of characteristic compounds specific to individual plant parts or vegetation stages was further addressed. The data revealed that classification of various vegetation phases based on quantitative evaluation of bufadienolides and dihydroquercetin derivatives applying principal component analysis (PCA) appears possible. Thus, the methodology presented here forms the basis for future routine application in quality control of raw materials and pharmaceutical preparations derived from sea squill. This will allow systematic comparison of different plant parts, vegetation stages and origins based on an extended sample set.

How do sink and source activities influence the reproduction and vegetative growth of spring ephemeral herbs under different light conditions?

Spring ephemeral herbs inhabiting deciduous forests commonly complete reproduction and vegetative growth before canopy closure in early summer. Effects of shading by early canopy closure on reproductive output and vegetative growth, however, may vary depending on the seasonal allocation patterns of photosynthetic products between current reproduction and storage for future growth in each species. To clarify the effects of sink-source balance on seed production and bulb growth in a spring ephemeral herb, Gagea lutea, we performed a bract removal treatment (source reduction) and a floral-bud removal treatment (sink reduction) under canopy and open conditions. Leaf carbon fixations did not differ between the forest and open sites and among treatments. Bract carbon fixations were also similar between sites but tended to decrease when floral buds were removed. Seed production was higher under open condition but decreased by the bract-removal treatment under both light conditions. In contrast, bulb growth was independent of light conditions and the bract-removal treatment but increased greatly by the bud-removal treatment. Therefore, leaves and bracts acted as specialized source organs for vegetative and reproductive functions, respectively, but photosynthetic products by bracts were flexibly used for bulb growth when plants failed to set fruits. Extension of bright period was advantageous for seed production (i.e., source limited) but not for vegetative growth (i.e., sink limited) in this species.

Somatic embryogenesis and synthetic seed production--a biotechnological approach for true-to-type propagation and in vitro conservation of an ornamental bulbaceous plant Drimiopsis kirkii Baker.

An efficient plant regeneration protocol through indirect somatic embryogenesis pathway via callus had been developed from the leaf explant of an ornamental bulbaceous plant Drimiopsis kirkii. Optimum friable calli were induced on Murashige and Skoog (MS) basal medium supplemented with 3.0 mg/l of 2,4-dichlorophenoxyacetic acid and 1.0 mg/l of α-naphthalene acetic acid (NAA). On subculturing the callus on MS medium supplemented with 2.5 mg/l of thidiazuron (TDZ), 73.3 % of the cultures responded with 20.4 ± 0.3 somatic embryos (SEs) per 500 mg callus at different stages of development after 6 weeks of culture. The highest response of 86.7 % with 28.3 ± 0.5 embryos per 500 mg callus was observed on MS medium supplemented with 2.5 mg/l TDZ and 1.0 mg/l NAA. SEs were encapsulated in calcium alginate beads for the production of synthetic seeds (SSs) and their storability was investigated. The highest SS germination (93.3 %) was observed in 1.0 % sodium alginate followed by 86.7 % germination with 2.5 % sodium alginate. The SSs were stored at three different temperatures (4, 15, and 24 ºC) up to 6 months. The SSs kept at 15 °C showed 64.4 % germinability even after 4 months of storage. Both nonencapsulated and encapsulated SE-derived plants were successfully transferred to soil with 93.3 and 88.3 % survival rate accordingly. Randomly amplified polymorphic DNA (RAPD) analysis revealed that there were no somaclonal variations among the plants produced via somatic embryogenesis and they are true-to-type to their parental plant. These results confirmed the most reliable methods, which can be further used for genetic transformation studies as well as for mass propagation of ornamental D. kirkii at a commercial level.

Carbon-nitrogen ratio and in vitro assimilate partitioning patterns in Cyrtanthus guthrieae L.

In response to variations in nutritional composition of the growth medium, plants often adjust their metabolism and progressively alter their growth patterns. Carbon (C) and nitrogen (N) constitute the major plant nutritional components influencing plant growth and development patterns. This study examined the growth dynamics and patterns of assimilate partitioning to primary and secondary metabolites in response to varying levels and combinations of C and N in the culture media of Cyrtanthus guthrieae. In vitro callus-derived C. guthrieae plantlets were cultured on solid Murashige and Skoog (MS) media with different concentrations and combinations of C and N. Relative growth rate (RGR) increased proportionally with an increase in C concentrations up to 88 mM sucrose (0.58 d(-1)) beyond which it was hardly influenced by further increases in C. Growth was also significantly favoured in media with high concentrations of N at all C concentrations tested. In C-limited media regimes with growth saturating N conditions, alkaloid accumulation became favoured while polyphenol content increased with an increase in C levels in the medium, a characteristic pattern that appeared to be less influenced by the amount of N. Of the primary metabolites, only proteins showed small significant variations across different media treatments, with starch and soluble sugars increasing proportionately with C levels. In the medium with a high sucrose concentration (175 mM), soluble sugars, amino acids and polyphenols increased markedly, possibly as an adaptive response to the reduced osmotic potential in the media and/or a storage mechanism for excess C and N reserves in the media. From a medicinal perspective, with regard to polyphenolic compounds in C. guthrieae, growth medium conditions that allow for high levels of C pools in the tissue would thus be favourable for the enhanced synthesis of this group of compounds. The medium conditions with 175 mM sucrose and 10.3 mM NH4NO3 gave the highest total polyphenols, flavonoids and proanthocyanidins with a moderate growth rate.

Overexpression of the gibberellin 2-oxidase gene from Torenia fournieri induces dwarf phenotypes in the liliaceous monocotyledon Tricyrtis sp.

Gibberellins (GAs) are the plant hormones that control many aspects of plant growth and development, including stem elongation. Genes encoding enzymes related to the GA biosynthetic and metabolic pathway have been isolated and characterized in many plant species. Gibberellin 2-oxidase (GA2ox) catalyzes bioactive GAs or their immediate precursors to inactive forms; therefore, playing a direct role in determining the levels of bioactive GAs. In the present study, we produced transgenic plants of the liliaceous monocotyledon Tricyrtis sp. overexpressing the GA2ox gene from the linderniaceous dicotyledon Torenia fournieri (TfGA2ox2). All six transgenic plants exhibited dwarf phenotypes, and they could be classified into two classes according to the degree of dwarfism: three plants were moderately dwarf and three were severely dwarf. All of the transgenic plants had small or no flowers, and smaller, rounder and darker green leaves. Quantitative real-time reverse transcription-polymerase chain reaction (PCR) analysis showed that the TfGA2ox2 expression level generally correlated with the degree of dwarfism. The endogenous levels of bioactive GAs, GA1 and GA4, largely decreased in transgenic plants as shown by liquid chromatography-mass spectrometry (LC-MS) analysis, and the level also correlated with the degree of dwarfism. Exogenous treatment of transgenic plants with gibberellic acid (GA3) resulted in an increased shoot length, indicating that the GA signaling pathway might normally function in transgenic plants. Thus, morphological changes in transgenic plants may result from a decrease in the endogenous levels of bioactive GAs. Finally, a possibility of molecular breeding for plant form alteration in liliaceous ornamental plants by genetically engineering the GA metabolic pathway is discussed.

[Study on the chromatographic fingerprints of Reineckia carnea from different habitats by HPLC-ELSD].

OBJECTIVE: To investigate the HPLC-ELSD fingerprints of Reineckia carnea from different habitats and provide a theoretical evidence to evaluate its quality. METHODS: The chromatographic conditions were Diamonsil (R) C18 column (4.6 mm x 250 mm, 5 microm) by linear gradient elution using water and acetonitrile as mobile phase at a flow rate of 0.8 mL/min, column temperature at 25 degrees C and sample injection 30 microL. An Agilent 1260 infintiy ELSD operated at 85 degrees C, gain 6 and 3.5 bar nitrogen. RESULTS: The HPLC-ELSD fingerprints were established with 6 common peaks and methodology reached the required standard. CONCLUSION: The method is simple, rapid, accurate, and can be used for quality evaluation of Reineckia carnea from different habituts.