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Background: Thymus vulgaris is a valuable medicinal plant widely cultivated for its aromatic and medicinal properties. However, like many plants, T. vulgaris faces challenges such as drought stress, which significantly affects its growth, morphological, physiological, and biochemical processes. Understanding how endophytic fungi isolated from Lamiaceae family influence T. vulgaris under varying watering regimes can enhance its resilience against drought stress. This study aims to assess the impact of individual and co-inoculation of three native endophytic species, i.e., Fusarium sp. (F 1 ), Cladosporium puyae (F 2 ), and Curvularia australiensis (F 3 ), on T. vulgaris growth parameters under different irrigation regimes in greenhouse conditions., Results: It has been discovered that using fungal endophytes as a biological tool can benefits T. vulgaris under drought stress. The results indicated that drought stress significantly reduced the growth, chlorophyll, and carotenoid content of plants lacking endophytes. Combinatory applications with fungal endophytes significantly improved the above-mentioned parameters under drought stress. Lipid peroxidation levels were significantly reduced in plants inoculated with bacterial endophytes. Drought stress significantly increased the activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), peroxidase (POD), and catalase (CAT) in drought conditions., Conclusions: The findings suggested that the addition of fungal endophytes to the inoculum enhances drought tolerance in T. vulgaris by mitigating the harmful impact of drought stress on plant growth and physiological functions. The higher activity of antioxidant enzymes and improved redox state of glutathione are responsible for plants' greater resistance to drought., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Glechoma longituba, recognized as a medicinal plant, provides valuable pharmaceutical raw materials for treating various diseases. Saline-alkali stress may effectively enhance the medicinal quality of G. longituba by promoting the synthesis of secondary metabolites. To investigate the changes in the primary medicinal components of G. longituba under saline-alkali stress and improve the quality of medicinal materials, Na 2 CO 3 was applied to induce short-term stress under different conditions and the biomass, physiologically active substances and primary medicinal components of G. longituba were measured in this study., Results: Under alkaline salt stress, the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) were elevated in G. longituba, accompanied by increased accumulation of proline (Pro) and malondialdehyde (MDA). Furthermore, analysis of the medicinal constituents revealed that G. longituba produced the highest levels of soluble sugars, flavonoids, ursolic acid, and oleanolic acid under 0.6% Na 2 CO 3 stress for 48 h, 0.2% Na 2 CO 3 stress for 72 h, 0.4% Na 2 CO 3 stress for 12 h, and 0.4% Na 2 CO 3 stress for 8 h, respectively., Conclusions: Short-term Na 2 CO 3 stress enhances the synthesis of medicinal components in G. longituba. By manipulating stress conditions, the production of various medicinal substances could be optimized. This approach may serve as a basis for the targeted cultivation of G. longituba, offering potential applications in the treatment of diverse diseases., (© 2024. The Author(s).)

AbstractPollen grains from different plants potentially compete for ovule access because flowers produce many more pollen grains than ovules. Pollen competition could occur on pollinators, where there is finite space for pollen placement. Here, we explore the explosive pollen deposition in Hypenia macrantha (Lamiaceae, a perennial flowering plant native to South America that is frequently visited by hummingbirds) and determine whether it can improve male performance by reducing pollen loads deposited by previously visited flowers. Through the simulation of floral visits utilizing a hummingbird skull, we showed that explosive pollen deposition by untriggered flowers dislodges almost twice as many pollen grains as already-triggered flowers. In addition, pollen removal increases with the amount of deposited pollen by the floral explosion, suggesting that the precision or the explosive force of pollen deposition plays a pivotal role in this pollen removal process. These results suggest that explosive pollen placement, a mechanism that has evolved in many unrelated angiosperm clades, may confer a prepollination male competition advantage to plants.

The distinctive characteristics of nanoparticles and their potential applications have been given considerable attention by scientists across different fields, particularly agriculture. However, there has been limited effort to assess the impact of copper nanoparticles (CuNPs) in modulating physiological and biochemical processes in response to salt-induced stress. This study aimed to synthesize CuNPs biologically using Solenostemma argel extract and determine their effects on morphophysiological parameters and antioxidant defense system of barley (Hordeum vulgare) under salt stress. The biosynthesized CuNPs were characterized by (UV-vis spectroscopy with Surface Plasmon Resonance at 320 nm, the crystalline nature of the formed NPs was verified via XRD, the FTIR recorded the presence of the functional groups, while TEM was confirmed the shape (spherical) and the sizes (9 to 18 nm) of biosynthesized CuNPs. Seeds of barley plants were grown in plastic pots and exposed to different levels of salt (0, 100 and 200 mM NaCl). Our findings revealed that the supplementation of CuNPs (0, 25 and 50 mg/L) to salinized barley significantly mitigate the negative impacts of salt stress and enhanced the plant growth-related parameters. High salinity level enhanced the oxidative damage by raising the concentrations of osmolytes (soluble protein, soluble sugar, and proline), malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). In addition, increasing the activities of enzymatic antioxidants, total phenol, and flavonoids. Interestingly, exposing CuNPs on salt-stressed plants enhanced the plant-growth characteristics, photosynthetic pigments, and gas exchange parameters. Furthermore, CuNPs counteracted oxidative damage by lowering the accumulation of osmolytes, H 2 O 2 , MDA, total phenol, and flavonoids, while simultaneously enhancing the activities of antioxidant enzymes. In conclusion, the application of biosynthesized CuNPs presents a promising approach and sustainable strategy to enhance plant resistance to salinity stress, surpassing conventional methods in terms of environmental balance., (© 2024. The Author(s).)

Background: Zataria multiflora Boiss. is a medicinal and aromatic plant from the Lamiaceae family. It is extensively used in Iranian traditional medicine, mostly as a replacement for Thyme species. This study was focused on the analysis of chemical composition and the distribution and types of trichomes of Z. multiflora grown under different conditions. Equilibrium headspace analysis in combination with GC-FID-MS was used to identify volatile compounds released by aerial parts of Z. multiflora in development stages of 50 and 100% flowering under normal and drought-stress conditions., Results: The main constituents were p-cymene (20.06-27.40%), γ-terpinene (12.44-16.93%), and α-pinene (6.91-16.58%) and thymol (8.52-9.99%). The highest content of p-cymene (27.40%) and thymol (9.99%) was observed in the 50% flowering stage at the 90% field capacity, while the maximum γ-terpinene (16.93%) content was recorded in the 100% flowering stage under normal conditions. Using the SEM method, it was found that peltate glandular and non-glandular trichomes are distributed on the surface of the leaf, stem, and outer side of the calyx. However, capitate trichomes only are detected on the stem and calyx in the 100% flowering and beginning of blooming stages, respectively. The type and structure of trichomes do not vary in different development stages, but they differ in density. The highest number of leaf peltate glandular trichomes was observed in the vegetative and beginning of blooming stages at 50% and 90% field capacity, respectively. Non-glandular trichomes of the stem were observed with high density in both normal and stress conditions, which are more densely in 90% field capacity., Conclusions: Since this plant has strong potential to be used in the food and pharmacological industries, this study provides valuable information for its cultivation and harvesting at specific phenological stages, depending on desired compounds and their concentrations., (© 2024. The Author(s).)

Plant species represent the hierarchical expression of vegetation as it is affected by various environmental gradients. We explored the plant species composition, distribution pattern, communities formation and their respective indicators under the influence of various environmental factors in the Dhirkot region, Azad Jammu and Kashmir. It was hypothesized that different environmental factors were responsible for the formation of various plant communities each with a distinct indicator. Quantitative ecological techniques were used for the sampling of vegetation. A total of 114 quadrats were established in 13 selected sampling sites. Phytosociological attributes were calculated for each plant species at each quadrat. Soil samples were collected and analyzed using different standard protocols. All the collected data were analyzed using Cluster Analysis, Indicator Species Analysis and Canonical Correspondence Analysis of PCORD and CANOCO software, respectively. A total of 145 plant species were recorded belong to 62 different families. Asteraceae and Lamiaceae were the dominant families, represented by 12 species each (8.27%). Cluster Analysis classify all the stations and plants into four major plant communities as 1) Olea-Desmodium-Prunilla community. 2) Abies-Zanthoxylum-Pteracanthus community 3) Cedrus-Elaeagnus-Hypericum community 4) Alnus-Myrsine-Ranunculus community. Soil pH, electrical conductivity, soil saturation, organic matter and altitude were the significant environmental factors that play its essential role in the plant species distribution, composition, formation of major plant communities and their respective indicators in the region. It is recommended that the identified indicator and rare plant species of the investigated area can further be grown for conservation and management purposes in in-situ environment., Competing Interests: The authors have declared that no competing interests exist.

The analysis of plant cell structure provides valuable information about its morphological, physiological, and biochemical characteristics. Nowadays, scanning electron microscope (SEM) is widely used to provide high-resolution images at the surface of biological samples. However, biological specimens require preparation, including dehydration and coating with conductive materials for imaging by SEM. There are several techniques for providing images with maximum maintenance of cell structure and minimum cellular damage, but each requires the use of expensive and hazardous materials, which can be damaging to the cell in many cases. Therefore, the provision of new and effective preparation methods based on maintaining cell structure for imaging can be very practical. In the present study, a fast and cost-effective protocol was first performed for chemical fixation and preparation of the plant cells for imaging by SEM. Taxus baccata and Zhumeria majdae cells were chemically fixed using glutaraldehyde and then successfully dried with different percentages of ethanol including 70, 80, 90, and 100%. In addition, SEM was performed for imaging the cell surface in different micro-scales. This protocol can be used by plant cell biologists and biotechnologists who are interested in studying structural and biochemical responses of treated or stressed plant cells by SEM., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Thidiazuron (TDZ) is a diphenylurea synthetic herbicide and plant growth regulator used to defoliate cotton crops and to induce regeneration of recalcitrant species in plant tissue culture. In vitro cultures of African violet thin petiole sections are an ideal model system for studies of TDZ-induced morphogenesis. TDZ induces de novo shoot organogenesis at low concentrations and somatic embryogenesis at higher concentrations of exposure. We used an untargeted metabolomics approach to identify metabolites in control and TDZ-treated tissues. Statistical analysis including metabolite clustering, pattern and pathway tools, logical algorithms, synthetic biotransformations and hormonomics identified TDZ-induced changes in metabolism. A total of 18,602 putative metabolites with extracted masses and predicted formulae were identified with 1412 features that were found only in TDZ-treated tissues and 312 that increased in response to TDZ. The monomer of TDZ was not detected intact in the tissues but putative oligomers were found in the database and we hypothesize that these may form by a Diels-Alder reaction. Accumulation oligomers in the tissue may act as a reservoir, slowly releasing the active TDZ monomer over time. Cleavage of the amide bridge released TDZ-metabolites into the tissues including organic nitrogen and sulfur containing compounds. Metabolomics data analysis generated six novel hypotheses that can be summarized as an overall increase in uptake of sugars from the culture media, increase in primary metabolism, redirection of terpene metabolism and mediation of stress metabolism via indoleamine and phenylpropanoid metabolism. Further research into the specific mechanisms hypothesized is likely to unravel the mode of action of TDZ and to provide new insights into the control of plant morphogenesis.

The aim of the study was to assess the effects of Melittis melissophyllum shading on its development and accumulation of phenolics. Their content (verbascoside, apiin, luteolin-7- O -glucoside, coumarin, 3,4-dihydroxycoumarin, o -coumaric acid 2- O -glucoside as well as o -coumaric, p -coumaric, chlorogenic, caffeic, ferulic and cichoric acid) was determined in the herb using HPLC-DAD. The results showed that the content of abovementioned flavonoids and phenolic acids was highest in plants grown under full sunlight. On the other hand, a higher content of coumarin was observed in shaded plants, especially after the seed-setting stage. A similar tendency was noted for the amount of chlorophyll a and b. The content of hydrogen peroxide and malondialdehyde, the activity of polyphenol oxidase and catalase and the antioxidant capacity of plant extracts (measured using DPPH, ABTS and FRAP assays) were found to be the highest in the plants grown in full sunlight. However, the plants grown in moderate (30%) shade were found to thrive best.

The incidence of fungal infections has been steadily increasing in recent years. Systemic mycoses are characterized by the highest mortality. At the same time, the frequency of infections caused by drug-resistant strains and new pathogens e.g., Candida auris increases. An alternative to medicines may be essential oils, which can have a broad antimicrobial spectrum. Rich in the essential oils are plants from the Lamiaceae family. In this review are presented antifungal activities of essential oils from 72 Lamiaceae plants. More than half of these have good activity (minimum inhibitory concentrations (MICs) < 1000 µg/mL) against fungi. The best activity (MICs < 100) have essential oils from some species of the genera Clinopodium , Lavandula , Mentha , Thymbra , and Thymus . In some cases were observed significant discrepancies between different studies. In the review are also shown the most important compounds of described essential oils. To the chemical components most commonly found as the main ingredients include β-caryophyllene (41 plants), linalool (27 plants), limonene (26), β-pinene (25), 1,8-cineole (22), carvacrol (21), α-pinene (21), p-cymene (20), γ-terpinene (20), and thymol (20).

Drought stress is an increasingly common and worrying phenomenon because it causes a loss of production in both agriculture and forestry. Teak is a tropical tree which needs alternating rainy and dry seasons to produce high-quality wood. However, a robust understanding about the physiological characteristics and genes related to drought stress in this species is lacking. Consequently, after applying moderate and severe drought stress to teak seedlings, an infrared gas analyzer (IRGA) was used to measure different parameters in the leaves. Additionally, using the root transcriptome allowed finding and analyzing the expression of several drought-related genes. As a result, in both water deficit treatments a reduction in photosynthesis, transpiration, stomatal conductance and leaf relative water content was found. As well, an increase in free proline levels and intrinsic water use efficiency was found when compared to the control treatment. Furthermore, 977 transcripts from the root contigs showed functional annotation related to drought stress, and of these, TgTPS1, TgDREB1, TgAREB1 and TgPIP1 were selected. The expression analysis of those genes along with TgHSP1, TgHSP2, TgHSP3 and TgBI (other stress-related genes) showed that with moderate treatment, TgTPS1, TgDREB1, TgAREB1, TgPIP1, TgHSP3 and TgBI genes had higher expression than the control treatment, but with severe treatment only TgTPS1 and TgDREB1 showed higher expression than the control treatment. At the end, a schematic model for the physiological and molecular strategies under drought stress in teak from this study is provided. In conclusion, these physiological and biochemical adjustments in leaves and genetic changes in roots under severe and prolonged water shortage situations can be a limiting factor for teak plantlets' growth. Further studies of those genes under different biotic and abiotic stress treatments are needed., Competing Interests: The authors have declared that no competing interests exist.

Transient thermal dissipation (TTD) systems provide a simple way to measure xylem sap flow with dual or single Granier-type probe, which gives lower energy consumption and higher accuracy due to its lessor sensitivity to thermal interferences. A new system, TTD heat within 5 min (TTD5), proposed on the reduction of the heating duration. This evolution captured interest through decreased energy consumption and increased temporal resolution. Within our study, the first objective was to test and calibrate this new system with a single probe for young rubber tree - Hevea brasiliensis. The second objective was to explore the sources of variability in calibration such as species, individual cut-stems and probe-wood contact. The complementary species consisted of two diffuse-porous species (mango tree - Mangifera indica, eucalyptus tree - Eucalyptus camaldulensis) and one ring-porous species (teak tree - Tectonia grandis). Twenty-eight response curves were assessed over a large range of flux densities from 0.5 to 10 l dm-2 h-1. The incremental rise of temperature from 30 to 300 s (T300-30) after commencement of heating was sensitive to flux density over the complete range. Compared with the full signal at 300 s, the incremental signal markedly reduced the variability between response curves within species and between species. Moreover, a new index K2, defined as (T0 - Tu)/T0, normalized the responses between 0 and 1. However, the responses had a non-linear trend above 5 l dm-2 h-1. Within diffuse-porous wood type, the species did not differ in calibration, whereas the ring-porous species was markedly different. A sigmoid function provided the best fit for the diffuse-porous species. Individual stems were identified as the main source of within-species variability in calibration. The normalizing K2 index removed the influence of probe-wood contacts, controlled through drilling difference; however, there was still an effect of individual stems interacting with flux density (P = 0.019). Replications of cut-stems and response curves are necessary to assess a reliable averaged calibration. In conclusion, the applicability of the TTD5 system with a single probe has been confirmed and several sources of variability in calibration have been evaluated., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Premise: Cambial activity in some tropical trees varies intra-annually, with the formation of xylem rings. Identification of the climatic factors that regulate cambial activity is important for understanding the growth of such species. We analyzed the relationship between climatic factors and cambial activity in four tropical hardwoods, Acacia mangium, Tectona grandis, Eucalyptus urophylla, and Neolamarckia cadamba in Yogyakarta, Java Island, Indonesia, which has a rainy season (November-June) and a dry season (July-October)., Methods: Small blocks containing phloem, cambium, and xylem were collected from main stems in January 2014, October 2015 and October 2016, and examined with light microscopy for cambial cell division, fusiform cambial cells, and expanding xylem cells as evidence of cambial activity., Results: During the rainy season, when precipitation was high, cambium was active. By contrast, during the dry season in 2015, when there was no precipitation, cambium was dormant. However, in October 2016, during the so-called dry season, cambium was active, cell division was conspicuous, and a new xylem ring formation was initiated. The difference in cambial activity appeared to be related to an unusual pattern of precipitation during the typically dry months, from July to October, in 2016., Conclusions: Our results indicate that low or absent precipitation for 3 to 4 months induces cessation of cambial activity and temporal periodicity of wood formation in the four species studied. By contrast, in the event of continuing precipitation, cambial activity in the same trees may continue throughout the year. The frequency pattern of precipitation appears to be an important determinant of wood formation in tropical trees., (© 2019 Botanical Society of America.)

Phylogenies recurrently demonstrate that oceanic island systems have been home to rapid clade diversification and adaptive radiations. The existence of adaptive radiations posits a central role of natural selection causing ecological divergence and speciation, and some plant radiations have been highlighted as paradigmatic examples of such radiations. However, neutral processes may also drive speciation during clade radiations, with ecological divergence occurring following speciation. Here, we document an exceptionally rapid and unique radiation of Lamiaceae within the New Caledonian biodiversity hotspot. Specifically, we investigated various biological, ecological, and geographical drivers of species diversification within the genus Oxera. We found that Oxera underwent an initial process of rapid cladogenesis likely triggered by a dramatic period of aridity during the early Pliocene. This early diversification of Oxera was associated with an important phase of ecological diversification triggered by significant shifts of pollination syndromes, dispersal modes, and life forms. Finally, recent diversification of Oxera appears to have been further driven by the interplay of allopatry and habitat shifts likely related to climatic oscillations. This suggests that Oxera could be regarded as an adaptive radiation at an early evolutionary stage that has been obscured by more recent joint habitat diversification and neutral geographical processes. Diversification within Oxera has perhaps been triggered by varied ecological and biological drivers acting in a leapfrog pattern, but geographic processes may have been an equally important driver. We suspect that strictly adaptive radiations may be rare in plants and that most events of rapid clade diversification may have involved a mixture of geographical and ecological divergence., (© The Author(s) 2018. Published by Oxford University Press on behalf of the Society of Systematic Biologists.)

Theory predicts that less fit individuals would disperse more often than fitter ones (Fitness Associated Dispersal, FAD hypothesis). To test this prediction under laboratory conditions, an entire life cycle of Lamium amplexicaule plants and the preferences of its dispersal agent, Messor ebeninus ants, were tracked. Characterization of individual L. amplexicaule plant revealed high variability in spot cover on the surface of the seeds, where less fit plants produce "unspotted seeds" (see Fig. 1 in Introduction). Unspotted L. amplexicaule seeds showed higher variation in germination time and lower germination rate. Moreover, M. ebeninus ants preferably collected these unspotted seeds. Our results show that low fitness L. amplexicaule plants produce seeds with higher potential for dispersal, supporting the FAD hypothesis in a plant-animal system.

The Central African Copperbelt of the DR Congo and Zambia hosts more than 30 known Cu-Co hyperaccumulator plant species. These plants can accumulate extraordinarily high concentrations of Cu and Co in their living tissues without showing any signs of toxicity. Haumaniastrum robertii is the most extreme Co hyperaccumulator (able to accumulate up to 1 wt% Co), whereas Aeolanthus biformifolius is the most extreme Cu hyperaccumulator (with up to 1 wt% Cu). The phenomenon of Cu-Co hyperaccumulator plants was studied intensively in the 1970s through to the 1990s, but doubts arose regarding earlier observations due to surficial contamination of plant material with mineral particles. This study set out to determine whether such extraneous contamination could be observed on herbarium specimens of Haumaniastrum robertii and Aeolanthus biformifolius using scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS). Further, synchrotron X-ray absorption spectroscopy (XAS) was used to identify the chemical forms of Cu and Co in newly collected Haumaniastrum katangense plant material from the DR Congo. The results show that surficial contamination is not the cause for abnormal Cu-Co concentrations in the plant material, but rather that Cu-Co enrichment is endogenous. The chemical form of Cu and Co (complexation with carboxylic acids) provides additional evidence that genuine hyperaccumulation, and not soil mineral contamination, is responsible for extreme tissue concentrations of Cu and Co in Haumaniastrum katangense.

Drought stress is one of the most important stresses adversely affecting plant growth and yield production. Due to the importance of global warming, the investigation of drought effects on the growth and quality of medical plants is of vital importance. Accordingly, a two-year field experiment was conducted in 2013-2014 to determine the effects of drought levels and plant species on Balangu (Lallemantia sp.) growth and physiological properties including medicinal content. The experiment was a split plot in which the drought levels (main plots) including control (D1, moisture field capacity, water potential at, 0.5 atm), moderate stress (D2, 6.5 atm) and severe stress (D3, 9.5 atm), and the Balangu species (sub plots) including Lallemantia royleana (Benth) (L1) and L. iberica (L2) were tested as the experimental treatments. Plant yield, oil content and the biochemical properties (i.e. medicinal content) including phenolic compounds, proline, carotenoids, and the activity of antioxidant enzymes including peroxidase (EC 1.11.1), super oxide dismutase (SOD, EC 1.15.1.1)) and ascorbate peroxidase (APX, EC 1.11.1.11) were determined. Drought stress significantly decreased crop yield and oil content. However, the production of phenolic compounds and proline as well as the activity of antioxidant enzymes, SOD and APX increased under stress. The species L2 was the more tolerant species under drought stress. The interesting point about this research work is the increased production of secondary metabolites (i.e. phenolic compounds) under stress, affecting both Balangu response and medicinal properties. Accordingly, it may be possible to regulate the production of secondary metabolites (medicinal contents) in Balangu species by adjusting the irrigating practices., (Copyright © 2018. Published by Elsevier Masson SAS.)

A new gall midge, Asphondylia nepetae sp. n. Viggiani (Diptera: Cecidomyiidae), causing flower gall on Clinopodium nepeta (L.) Kuntze (Lamiaceae), is described from Europe. The morphological characteristics of adult, larvae, and pupa are described and illustrated. Molecular approach (by sequencing 28S-D2, ITS2, and COI) confirmed that A. nepetae is a distinct species. The development of the gall is always associated with the presence of the fungus Botryosphaeria dothidea (Moug.: Fr.) Ces. and De Not. (Botryosphaeriales: Botryosphaeriaceae). The new species can complete several generations per year, on the flowers of the same host plant and its adults emerge from late spring to autumn. Pupae overwinter inside peculiar flower galls in a state of quiescence. The impact of the pest is highly variable with a percentage of flowers infested that ranged between 3 and 57.5% in the sampled years. Insect mortality was, at least in part, due to parasitoids that attack the young stages of the midge. Among them, the dominant species was Sigmophora brevicornis (Panzer) (Chalcidoidea: Eulophidae).

Ecological differentiation and genetic isolation are thought to be critical in facilitating coexistence between related species, but the relative importance of these phenomena and the interactions between them are not well understood. Here, we examine divergence in abiotic habitat affinity and the extent of hybridization and introgression between two rare species of Monardella (Lamiaceae) that are both restricted to the same serpentine soil exposure in California. Although broadly sympatric, they are found in microhabitats that differ consistently in soil chemistry, slope, rockiness and vegetation. We identify one active hybrid zone at a site with intermediate soil and above-ground characteristics, and we document admixture patterns indicative of extensive and asymmetric introgression from one species into the other. We find that genetic distance among heterospecific populations is related to geographic distance, such that the extent of apparent introgression is partly explained by the spatial proximity to the hybrid zone. Our work shows that plant species can maintain morphological and ecological integrity in the face of weak genetic isolation, intermediate habitats can facilitate the establishment of hybrids, and that the degree of apparent introgression a population experiences is related to its geographic location rather than its local habitat characteristics., (© 2018 John Wiley & Sons Ltd.)

Background and Objective: Flower resources are prerequisites for survival and reproduction of aphidophagous hoverflies adults. It is, therefore, necessary to evaluate their foraging behavior with regards to different flowering species in order to utilize hoverflies to enhance conservation biological control (CBC) of aphids., Materials and Methods: The foraging behavior of the female hoverfly, Sphaerophoria macrogaster (Thomson) was observed in coriander (Coriandrum sativum Linnaeus) and blue salvia (Salvia farinacea Bentham) flower patches using a focal sampling method combined with continuous recording. The sequences and durations of all foraging bouts were recorded during the residence of observed S. macrogaster in each flower patch., Results: Significantly more transitions from searching to assessment of flowers were noted in coriander than in blue salvia flower patches (χ2 = 4.55, p<0.05). However, approaching to probing transitions were significantly more frequent in blue salvia than in coriander flower patches (χ2 = 9.59, p<0.05). Foragers showed significantly prolonged inter plant movement but shorter probing durations in coriander flower patches. Of interest, total duration of time spent in patches by S. macrogaster did not differ significantly between coriander and blue salvia., Conclusion: Results from the present study illustrated that both flowering plant species were apparently attractive to Sphaerophoria macrogaster females but these foragers preferred coriander flowers. This finding may form the basis of flower selection in and around the crop fields for enhancement of this aphidophagous natural enemy to maximize the biological control of aphids.

Closely associated microbes have been shown to drive local adaptation of plants. However, few studies provide direct evidence, disclosing the role of arbuscular mycorrhiza fungi (AMF) in their rapid adaptation of plants toward heavy metal tolerance. Elsholtzia splendens is a Cu-tolerant plant that was used as a model plant to study seed morphological traits as well as traits related to seed germination and seedling growth. This was achieved after acclimation for two generations with 1000mg/kg CuSO 4 in either absence or presence of AMF. In the absence of AMF, acclimation to Cu for two generations significantly decreased surface area, perimeter length, and perimeter width of E. splendens seeds, as well as seedling survival rate and fresh weight of the radicle of seedlings. However, in the presence of AMF, both the germination rate and the germination index of E. splendens seeds as well as the fresh weights of hypocotyl and radicle significantly increased. These results revealed that after Cu acclimation treatment, seeds and seedlings that had been inoculated with AMF outperformed those without AMF inoculation under Cu addition, indicating that AMF can facilitate rapid adaptation of E. splendens to Cu stress. In addition, two generations of Cu acclimation under AMF absence significantly increased radicle length, while amplitude increased under AMF presence, indicating that the direct adaptive plasticity response of radicle length to Cu stress helps with the Cu stress adaptation of E. splendens., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Premise of the Study: The subtribe Menthinae (Lamiaceae), with 35 genera and 750 species, is among the largest and most economically important subtribes within the mint family. Most genera of Menthinae are found exclusively in the New World, where the group has a virtually continuous distribution ranging from temperate North America to southern South America. In this study, we explored the presence, timing, and origin of amphitropical disjuncts within Menthinae., Methods: Our analyses were based on a data set consisting of 89 taxa and the nuclear ribosomal DNA markers ITS and ETS. Phylogenetic relationships were determined under maximum likelihood and Bayesian criteria, divergence times were estimated with the program BEAST, and ancestral range estimated with BioGeoBEARS., Key Results: A North Atlantic Land Bridge migration event at about 10.6 Ma is inferred from western Eurasia to North America. New World Menthinae spread rapidly across North America, and then into Central and South America. Several of the large speciose genera are not monophyletic with nuclear rDNA, a finding mirrored with previous chloroplast DNA results. Three amphitropical disjunctions involving North and southern South America clades, one including a southeastern South American clade with several genera, were inferred to have occurred within the past 5 Myr., Conclusions: Although three New World Menthinae genera occur in both North and South America, none exhibit an amphitropical disjunction. However, three clades exhibit amphitropical disjunctions, all dating to the early Pliocene, and all involve jump dispersals to either southeastern or southwestern South America from southeastern North America., (© 2017 Botanical Society of America.)

The diverse pigmentation patterns of flower corollas probably result from pollinator-mediated selection. Previous studies demonstrated that R2R3-MYB factors may have been recruited in the regulation of corolla pigmentation. However, how R2R3-MYBs became so diverse in their regulation of different pigmentation patterns remains unclear. Here, we studied a Lamiales species, Torenia fournieri, which has elaborate zygomorphic flowers with dorsal-ventral asymmetries in corolla pigmentation. We found recent gene duplication events in CYCLOIDEA-like (CYC-like) and RADIALIS-like (RAD-like) genes, and functionally analyzed three dorsal-specific expression factors: TfCYC1, TfCYC2, and TfRAD1. We found that the CYC-RAD module coordinates petal shape and corolla pigmentation, as ectopic expression of TfCYC2 or TfRAD1 disrupted the asymmetric corolla pigmentation pattern and produced strongly dorsalized flowers. Dorsal petal identity was lost when TfCYC2 was down-regulated or when TfRAD1 was knocked out. In T. fournieri, the diversified CYC and RAD genes have evolved regulatory loops, and TfCYC2 binds directly to the regulatory regions of an R2R3-MYB factor gene, TfMYB1, which might lead to its asymmetric expression and ultimately establish the asymmetric pigmentation pattern. These findings support the existence of a regulatory module that integrates dorsal-ventral patterning and asymmetric corolla pigmentation in T. fournieri., (© 2017 Sun Yat-sen University New Phytologist © 2017 New Phytologist Trust.)

Exposure to high salinity can trigger acclimation in many plants. Such an adaptative response is greatly advantageous for plants and involves extensive reprogramming at the molecular level. Acclimation allows plants to survive in environments that are prone to increasing salinity. In this study, diploid and autotetraploid Paulownia fortunei seedlings were used to detect alterations in leaf proteins in plants under salt stress. Up to 152 differentially abundant proteins were identified by Multiplex run iTRAQ-based quantitative proteomic and LC-MS/MS methods. Bioinformatics analysis suggested that P. fortunei leaves reacted to salt stress through a combination of common responses, such as induced metabolism, signal transduction, and regulation of transcription. This study offers a better understanding of the mechanisms of salt tolerance in P. fortunei and provides a list of potential target genes that could be engineered for salt acclimation in plants, especially trees.

Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. However, critical and noncritical structures in the disaccharide have not been dissected deeply. Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity., (© 2017 American Society of Plant Biologists. All Rights Reserved.)

Extrafloral structures are supposed to have evolved to protect flowers from harsh physical environments but might have effects on pollination. Overlapping leaves cover flowers in Eriophyton wallichii, an alpine perennial endemic to the Himalaya-Hengduan Mountains. In previous study, it has showed that these extrafloral leaves can protect interior flowers from temperature fluctuations caused by drastic solar radiation fluctuations, but these leaves may also protect interior flowers from rain wash and UVB damage, and we do not know which one is the main function. In this study, we investigated whether rain and UVB protection are the main functions of overlapping leaves covering flowers and their potential impact on pollination. We first measured the intensities of UVB radiation in open air, beneath leaves and corollas, and then examined pollen susceptibility to different intensities of UVB and rain in the laboratory to estimate whether corollas per se protect interior pollen from UVB and rain damage. We also carried out pollination treatments and observed pollinator visitation of flowers with and without leaves in the field to assess whether the overlapping leaves covering flowers impair pollinator attraction. Our results showed that (1) water and strong UVB significantly decreased pollen germinability, but corollas per se could protect pollen from UVB and rain damage; (2) no autonomous self-pollination and apomixis occurred, and pollinators were essential for the reproduction of E. wallichii; however, flower coverage by overlapping leaves did not limit pollination. We suggested that rain and UVB protection was not the main function of overlapping leaves covered flowers, given that this protection can be provided by corollas per se. Alternatively, this extrafloral structure in E. wallichii may have evolved in response to extreme high temperatures associated with the strong solar radiation fluctuations. This indicates that, even in alpine plants, extreme high temperature may affect the evolution of plant extrafloral structures., Competing Interests: The authors have declared that no competing interests exist.

Facilitation, or positive plant-plant interaction, has received increasing concern from ecologists over the last two decades. Facilitation may occur through direct mitigation of severe environments or indirect mediation by a third participant from the same or different trophic levels. The copper (Cu) tolerant species Elsholtzia splendens facilitates the establishment and growth of co-occurring Commelina communis through indirect enrichment of microbial activity. However, whether and how E. splendens impacts the microbial community that is associated with C. communis is less known. We characterized the soil bacterial community in the rhizosphere of C. communis in the absence and presence of E. splendens using PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and sequencing. The result showed that the richness of the bacterial community increased, but diversity and evenness remained similar, in the presence of E. splendens. Chloroflexi, Acidobacteria and Proteobacteria were the most dominant bacteria. The relative abundance of dominant and minor bacterial groups showed distinctly different responses to E. splendens. Principal component analysis and redundancy analysis indicated that variation of the bacterial community was determined by multiple factors and might be driven by the tested soil parameters collectively, or alternatively changed through plant root exudates or other microorganisms. Our results enhance the understanding of how the bacterial community associated with a beneficiary plant responds to a benefactor plant and suggests that the changes of bacterial community composition may have far-reaching influence on plant-soil feedback and the aboveground plant community in the long run., (Copyright © 2016. Published by Elsevier B.V.)

Gynodioecy, a sexual system where females and hermaphrodites co-occur, is found in << 1% of angiosperm species. To understand why gynodioecy is rare, we need to understand why females are maintained in some lineages, but not in others. We modelled the evolution of gynodioecy in the Lamiaceae, and investigated whether transition rates between gynodioecious and nongynodioecious states varied across the family. We also investigated whether the evolution of gynodioecy was correlated with the evolution of a herbaceous growth form and temperate distribution. Transition rates differed between Lamiaceae subfamilies. In the Nepetoideae, there were many transitions towards gynodioecy (n = 11), but also many reversions to nongynodioecy (n = 29). In addition, a herbaceous growth form, but not a temperate distribution, affected the rate of transitions both towards and away from gynodioecy; transitions towards gynodioecy occurred ˜16 times more frequently and transitions away from gynodioecy occurred ˜11 times more frequently in herbaceous lineages than in woody lineages. Within the Lamiaceae, lineages in which gynodioecy has frequently evolved also have a high rate of reversions to the nongynodioecious state. Consequently, to understand why gynodioecy is rare, we need to understand why sexual systems are more evolutionarily labile in some lineages than in others., (© 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.)

A tropical small tree, Callicarpa saccata, is known to have a symbiotic relationship with ants. It has sac-like structures at the base of the leaves that are inhabited by ants. No other species has been determined to be a myrmecophyte among the ca. 140 species of this genus. However, our recent field investigation discovered that two other species on Borneo (C. barbata and C. teneriflora) have hollow stems, which seem to be inhabited by ants. We observed the morphological features of these species in relation to their usage by ants, and became convinced that they are mymecophytic species. The molecular phylogenetic analyses using ITS and chloroplast regions suggest that C. saccata and C. teneriflora are closely related, but the differences in the myrmecophytic features of these species should be noted.

Some plants secrete toxic nectar to appeal to most effective pollinators and deter non-pollinators or nectar thieves; however available information about ecological function of toxic nectar remains scarce. Elsholtzia rugulosa stands out as a plant with toxic nectar recorded in SW China. We focused on the functional significance of the phenolic compound that imparts toxic to the nectar of E. rugulosa. The effects of phenolic nectar were studied in three visitors of the flowers of the winter-blooming E. rugulosa Hemsl. (Lamiaceae) in SW China. The pollinating species Apis cerana Fabricius (Apidae; Asian honey bee) and two occasional visitors, Vespa velutina Lepeletier (Vespidae; yellow-legged Asian hornet) and Bombus eximius Smith (Apidae; a bumble bee) were tested for their preferences for low and high concentrations of 4-hydroxybenzoic acid in hexose and sucrose solutions. The pollinator is important for the plant, which is dependent on pollinator visits to attain a higher seed production and it is most likely that the combination of phenolic toxic nectar and the adaptation to phenolic nectar by A. cerana delivers an evolutionary advantage to both actors. The low and high concentrations of the phenolic acid were nearly totally refused by both occasional visitors V. velutina and B. eximius and were preferred by the pollinator A. cerana. E. rugulosa gains by having a much higher seed production and the pollinating honey bee by having an exclusive and reliable food source during the winter season at high altitudes in SW China. We found that the function of the toxic phenolic compound has dual roles by appealing to legitimate pollinators and deterring non-pollinators of E. rugulosa.

Planting floral margins in agricultural landscapes has been shown to increase the abundance of pollinators in agro-ecosystems. However, to create efficient margins, it is necessary to use attractive, not weedy native plants with different blooming periods to prolong the availability of floral resources. Six native perennial plants of the Lamiaceae with different blooming periods were studied in a randomized block design, with the final aim to select the most efficient plants in floral mixtures by studying relationships between their floral phenology, floral density, and attractiveness to pollinators in Central Spain. In addition, their spatial expansion, i.e., potential weediness, was estimated under the field conditions, as the final purpose of the plants is to be implemented within the agro-ecosystems. The results showed that plant species with higher floral density (Nepeta tuberosa L. and Hyssopus officinalis L.) showed significantly higher attractiveness to pollinators and enhanced the attractiveness of floral mixtures. Species that bloomed in early spring (Salvia verbenaca L.) and in summer (Melissa officinalis L. and Thymbra capitata L.) did not efficiently contribute to the attractiveness of the mixtures to pollinators. In addition, besides high floral density of Salvia officinalis L. and N. tuberosa in the spring, warm and dry weather in spring 2012 enhanced the activity of bees, while cold and rainy weather in spring 2013 enhanced the activity of hoverflies. None of the plants showed weedy growth and so posed no danger of invading adjacent crops.

Plant responses to heavy metal contamination may depend on the presence of arbuscular mycorrhizal fungi (AMF). Elsholtzia splendens is an indicator species for the presence of copper (Cu) mines because both its flowering phenology and reproduction are tolerant to heavy metals. To test whether effects of Cu on the flowering phenology and reproduction of E. splendens depend on the presence of AMF, we conducted a factorial experiment with two Cu treatments (with or without Cu addition) crossed with two AMF treatments (with or without AMF inoculation). Without AMF, Cu addition significantly delayed the onset dates, ending dates and peak dates of flowering and decreased flowering duration. However, AMF inoculation reversed the effects of Cu stress, with recovered flowering onset and ending dates and increased the flowering duration. Cu addition significantly decreased inflorescence width and number, inflorescence biomass, vegetative biomass and total seed number, but significantly increased 1000-seed weight. AMF inoculation significantly increased vegetative biomass. Two-way ANOVA results showed that the interactive effects between Cu addition and AMF inoculation were significant on the inflorescence number, vegetative biomass and total seed number. These results indicate that AMF can alleviate the Cu stress on the flowering phenology and reproduction of E. splendens.

Objective: To investigate the phenotypic trait variation range of Clerodendranthus spicatus, and to look for phenotypic traits closely related with its yield and quality, in order to provide reference for its breeding., Methods: Randomly labelled plants of Clerodendranthus spicatus, observed its phenotypic traits and analyzed by variation, principal component, correlation and path analysis., Results: 13 phenotypic traits in the 15 germplasms of Clerodendranthus spicatus had great variations, the variations mainly distributed in yield, growth and genetic characteristics. Correlation and path analysis showed that, the plant dry weight had an extremely significantly positive correlation with fresh weight, and a positive correlation with stem height, stem diameter and root diameter. Plant fresh weight had a majorly direct contribution to the plant dry weight, stem height, stem diameter and root diameter also had a direct contribution to the plant dry weight. The other characters, including root length, branches, the number of leaf nodes, leaf number, leaf length, leaf width, fresh weight/dry weight ratio, rosmarinic acid content and ursolic acid content all had a negatively direct contribution to the plant dry weight. Rosmarinic acid content had a positive correlation with fresh weight, and a significantly positive correlation with fresh weight/dry weight ratio. Fresh weight had a majorly direct contribution to the rosmarinic acid content, stem height and stem diameter also had a direct contribution to the plant rosmarinic acid content. The other characters, including root length, root diameter, branches, the number of leaf nodes, leaf length, leaf width, dry weight, fresh weight/dry weight ratio, and ursolic acid content all had a negatively direct contribution to the rosmarinic acid content., Conclusion: The phenotypic traits of Clerodendranthus spicatus had rich variations on yield, growth and genetic characteristics. When choosing good germplasm, plant fresh weight, stem height, stem diameter and plant fresh weight/dry weight ratio having promoting effects on the plant dry weight and rosmarinic acid content should be considered comprehensively, and plant with vigorous growth, tall stem and more leaves can be choosed firstly.

The influence of locally different species interactions on trait evolution is a focus of recent evolutionary studies. However, few studies have demonstrated that geographically different pollinator-mediated selection influences geographic variation in floral traits, especially across a narrow geographic range. Here, we hypothesized that floral size variation in the Japanese herb Prunella vulgaris L. (Lamiaceae) is affected by geographically different pollinator sizes reflecting different pollinator assemblages. To evaluate this hypothesis, we posed two questions. (1) Is there a positive correlation between floral length and the proboscis length of pollinators (bumblebees) across altitude in a mountain range? (2) Does the flower-pollinator size match influence female and male plant fitness? We found geographic variation in the assemblage of pollinators of P. vulgaris along an altitudinal gradient, and, as a consequence, the mean pollinator proboscis length also changed altitudinally. The floral corolla length of P. vulgaris also varied along an altitudinal gradient, and this variation strongly correlated with the local pollinator size but did not correlate with altitude itself. Furthermore, we found that the size match between the floral corolla length and bee proboscis length affected female and male plant fitness and the optimal size match (associated with peak fitness) was similar for the female and male fitness. Collectively, these results suggest that pollinator-mediated selection influences spatial variation in the size of P. vulgaris flowers at a fine spatial scale., (© 2015 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2015 European Society For Evolutionary Biology.)

Differences in copper (Cu) absorption and transport, physiological responses and structural characteristics between two types of Cu-resistant plants, Oenothera glazioviana (Cu-exclusion type) and Elsholtzia haichowensis (Cu-enrichment type), were investigated in the present study. The results indicated the following: (1) After 50 μM Cu treatment, the Cu ratio in the xylem vessels of E. haichowensis increased by 60%. A Cu adsorption experiment indicated that O. glazioviana exhibited greater resistance to Cu, and Cu absorption and the shoot/root ratio of Cu were significantly lower in O. glazioviana than in E. haichowensis. (2) An analysis of the endogenous abscisic acid (ABA) variance and exogenous ABA treatment demonstrated that the ABA levels of both plants did not differ; exogenous ABA treatment clearly reduced Cu accumulation in both plants. (3) The leaf stomatal density of O. glazioviana was significantly less than that of E. haichowensis. Guard cells in E. haichowensis plants were covered with a thick cuticle layer, the epidermal hair was more numerous and longer, and the number of xylem conduits in the root was small. (4) The transpiration rate and the stomatal conductance of O. glazioviana were both significantly lower than those of E. haichowensis, regardless of whether the plants were treated with Cu. Taken together, these results indicate that the differences in the structural characteristics between these two plant species, particularly in the characteristics related to plant transpiration, are important factors that govern whether plants acquire or exclude Cu.

Seed dispersal by ants (myrmecochory) is found worldwide, but the benefits that plants obtain from this mutualism remain uncertain. In the present study, we conducted laboratory experiments to demonstrate seed predator avoidance as a benefit of myrmecochory using the annual ant-dispersed herb Lamium amplexicaule, the disperser ant Tetramorium tsushimae, and the seed predatory burrower bug Adomerus rotundus. We compared the predation intensity of Lamium amplexicaule seeds by Adomerus rotundus under the presence or absence of Tetramorium tsushimae. Both the number of seeds sucked by Adomerus rotundus adults and the feeding duration of sucked seeds by nymphs were significantly reduced in the presence of ants. This effect was most likely due to the behavioral alteration of Adomerus rotundus in response to the ant presence, because ants seldom predated Adomerus rotundus during the experiment. Our results demonstrated that the presence of ants decreases post-dispersal seed predation, even when the ants do not bury the seeds. The present study thus suggests that the non-consumptive effects of ants on seed predators benefit myrmecochorous plants.

Main Conclusion: We found the novel role of hydrogen sulfide in the adaptation of the alpine plant to altitude gradient in the Northern Tibetan Plateau. Alpine plants have developed strategies to survive the extremely cold conditions prevailing at high altitudes; however, the mechanism underlying the evolution of these strategies remains unknown. Hydrogen sulfide (H2S) is an essential messenger that enhances plant tolerance to environmental stress; however, its role in alpine plant adaptation to environmental stress has not been reported until now. In this work, we conducted a comparative proteomics analysis to investigate the dynamic patterns of protein expression in Lamiophlomis rotata plants grown at three different altitudes. We identified and annotated 83 differentially expressed proteins. We found that the levels and enzyme activities of proteins involved in H2S biosynthesis markedly increased at higher altitudes, and that H2S accumulation increased. Exogenous H2S application increased antioxidant enzyme activity, which reduced ROS (reactive oxygen species) damage, and GSNOR (S-nitrosoglutathione reductase) activity, which reduced RNS (reactive nitrogen species) damage, and activated the downstream defense response, resulting in protein degradation and proline and sugar accumulation. However, such defense responses could be reversed by applying H2S biosynthesis inhibitors. Based on these findings, we conclude that L. rotata uses multiple strategies to adapt to the alpine stress environment and that H2S plays a central role during this process.

Hydrogen sulfide (H2S) is an important gaseous transmitter in organisims. It widespreads in the organs and tissues of animals and participates in the biological process of cardiovascular relaxation, cell apoptosis and protection, inflammation and neuromodulation. H2S also can be synthesized in plants system and is involved in stress responses and the biological process of growth and development. This review describes the synthesis and biological function of H2S in plants. Based on our research for the adaptation of Lamiophlomis rotata to different altitude gradients, we firstly proposed H2S plays an important role in the adaptation of Lamiophlomis rotata to alpine environment.

Background and Aims: Copper (Cu) is an essential micronutrient for plants. However, excess amounts of Cu are toxic and result in a wide range of harmful effects on the physiological and biochemical processes of plants. Cell wall has a crucial role in plant defense response to toxic metals. To date, the process of cell wall response to Cu and the detoxification mechanism have not been well documented at the proteomic level., Methods: An recently developed 6-plex Tandem Mass Tag was used for relative and absolute quantitation methods to achieve a comprehensive understanding of Cu tolerance/detoxification molecular mechanisms in the cell wall. LC-MS/MS approach was performed to analyze the Cu-responsive cell wall proteins and polysaccharides., Key Results: The majority of the 22 up-regulated proteins were involved in the antioxidant defense pathway, cell wall polysaccharide remodeling, and cell metabolism process. Changes in polysaccharide amount, composition, and distribution could offer more binding sites for Cu ions. The 33 down-regulated proteins were involved in the signal pathway, energy, and protein synthesis., Conclusions: Based on the abundant changes in proteins and polysaccharides, and their putative functions, a possible protein interaction network can provide new insights into Cu stress response in root cell wall. Cu can facilitate further functional research on target proteins associated with metal response in the cell wall.

The white dead nettle, Lamium album L., is an herb that has been successfully cultivated under in vitro conditions. The L. album micropropagation system offers a combination of factors (light intensity, temperature, carbon dioxide (CO2) level, humidity) that are limiting for plant growth and bioactive capacity. To get a better understanding of the mechanism of plant acclimation towards environmental changes, we performed a comparative investigation on primary and secondary metabolism in fully expanded L. album leaves during the consecutive growth in in situ, in vitro, and ex vitro conditions. Although the genetic identity was not affected, structural and physiological deviations were observed, and the level of bioactive compounds was modified. During in vitro cultivation, the L. album leaves became thinner with unaffected overall leaf organization, but with a reduced number of palisade mesophyll layers. Structural deviation of the thylakoid membrane system was detected. In addition, the photosystem 2 (PS2) electron transport was retarded, and the plants were more vulnerable to light damage as indicated by the decreased photoprotection ability estimated by fluorescence parameters. The related CO2 assimilation and transpiration rates were subsequently reduced, as were the content of essential oils and phenolics. Transfer of the plants ex vitro did not increase the number of palisade numbers, but the chloroplast structure and PS2 functionality were recovered. Strikingly, the rates of CO2 assimilation and transpiration were increased compared to in situ control plants. While the phenolics content reached normal levels during ex vitro growth, the essential oils remained low. Overall, our study broadens the understanding about the nature of plant responses towards environmental conditions., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Background: Gesneriaceae family contains numerous species endemic to China, and many of them are listed as endangered species. There is a need for a simple and efficient method for long-term conservation of these species., Objective: The study aimed to establish an efficient procedure for cryopreserving Paraisometrum mileense, a critically endangered species endemic to Yunnan, China., Methods: Effects of sucrose concentration of preculture solution, duration of sucrose preculture, duration of plant vitrification solution 2 (PVS2) treatment, and cold acclimation on regeneration of cryopreserved adventitious shoot tips (ASTs) were assessed., Results and Conclusion: Among different sucrose preculture regimes tested, preculture with 0.3M sucrose for 24h resulted in best regeneration of cryopreserved ASTs. PVS2 treatment also affected regeneration considerably with the maximum survival of ASTs after incubation in PVS2 for 90 min at 0 degrees C. With the optimised parameters, the level of shoot regeneration from cryopreserved ASTs reached 86%. No morphological abnormalities were observed during one year's growth of the plantlets developing from cryopreserved ASTs. Procedure established in this research is a promising technique for the cryopreservation of ASTs of this species.

In this study, a hydroponics experiment was conducted to investigate the characteristics of Cd tolerance and accumulation of Elsholtzia argyi natively growing on the soil with high levels of heavy metals in a Zn/Pb mining site. Seedlings of E. argyi grown for 4 weeks and then were treated with 0(CK), 5,10,15, 20, 25, 30, 40, 50,100 umM Cd for 21 days. Each treatment had three replications. No visual toxic symptoms on shoots of E. argyi were observed at Cd level < or = 50 muM. The results indicated that the dry biomass of each tissue and the whole plants of the treatments with < or =40 umM cadmium were similar to that of the control, implying that E. argyi was a cadmium tolerant plant. The results also showed that the shoot Cd concentration significantly (P < 0.05) increased with the increase in the Cd level in nutrient solution. The shoot Cd concentration of the treatment with 40 umM Cd was as high as 237.9 mg kg(-1), which was higher than 100 mg kg(-1), normally used as the threshold concentration for identifying the Cd hyperaccumulating plant. It could be concluded that E. argyi was a Cd tolerant and accumulating plant species.

The present study aimed to test a hypothesis that acid invertases in root of metallophytes might play important roles in root growth under heavy metal stress. Plants of two contrasting populations, one from an ancient Cu mine (MP) and the other from a non-contaminated site (NMP), of metallophyte Elsholtzia haichowensis were treated with Cu in controlled experiments. The results showed that MP was Cu tolerant under 10 μM Cu2+ treatment. Cu treatment resulted in a higher root/shoot biomass ratio in MP compared to NMP. Scaling exponent in root/shoot allometric function in MP was lower than NMP. More complicated root architecture was observed in MP under Cu stress. Four full-length cDNAs (EhNcwINV, EhCcwINV, EhNvINV and EhCvINV) encoding cell wall and vacuolar invertases were cloned. Both of the transcript level and activity of the acid invertase in MP elevated under Cu treatment. There were positive correlations between root acid invertase transcript level, activity and root/shoot biomass ratio. The results indicated important roles of acid invertase in governing root growth under Cu stress. It also suggested that there was a possible interrelation between acid invertases and Cu tolerance mechanisms in MP of E. haichowensis.

Background and Aims: Gloxinia (Sinningia speciosa) is a popular commercial plant for its attractive and colourful flowers. However, the genetic mechanism of flowering time regulation in gloxinia is largely unknown. Recent studies on model plants have elucidated that miR159 acts as a negative regulator of floral transition in short-day photoperiods. The aim of this study was to investigate whether genetic modification of miR159 expression can offer an effective approach for regulation of flowering characteristics in gloxinia., Methods: Transgenic gloxinia plants were generated that over-express or suppress miR159 by means an efficient Agrobacterium-mediated transformation system in order to study the effect of miR159 on flowering time. In addition, the full-length cDNA of gloxinia GAMYB (SsGAMYB) was also cloned, and was verified to be a target of miR159 by modified RNA ligase-mediated 5' rapid amplification of cDNA ends., Key Results: Transgenic gloxinia plants that over-express or suppress miR159 exhibited significantly late or early flowering, respectively. During flower development, the expression level of miR159 was negatively correlated with SsGAMYB in gloxinia. MiR159-mediated SsGAMYB expression affected the expression levels of SsLEAFY (SsLFY) and three MADS-box genes (SsAP1, SsAP3 and SsAG), which regulated floral transition downstream of GAMYB. In addition, suppression of miR159 caused a conversion of petals and sepals in a few transgenic plants., Conclusions: miR159-regulated GAMYB expression is an effective pathway of flowering time control in gloxinia. Transgenic manipulation of miR159 can be used as an applicable strategy to regulate flowering time in commercial ornamental plants.

Background and Aims: There is a great need to search for natural compounds with superior prebiotic, antioxidant and immunostimulatory properties for use in (food) applications. Raffinose family oligosaccharides (RFOs) show such properties. Moreover, they contribute to stress tolerance in plants, acting as putative membrane stabilizers, antioxidants and signalling agents., Methods: A large-scale soluble carbohydrate screening was performed within the plant kingdom. An unknown compound accumulated to a high extent in early-spring red deadnettle (Lamium purpureum) but not in other RFO plants. The compound was purified and its structure was unravelled with NMR. Organs and organ parts of red deadnettle were carefully dissected and analysed for soluble sugars. Phloem sap content was analysed by a common EDTA-based method., Key Results: Early-spring red deadnettle stems and roots accumulate high concentrations of the reducing trisaccharide manninotriose (Galα1,6Galα1,6Glc), a derivative of the non-reducing RFO stachyose (Galα1,6Galα1,6Glcα1,2βFru). Detailed soluble carbohydrate analyses on dissected stem and leaf sections, together with phloem sap analyses, strongly suggest that stachyose is the main transport compound, but extensive hydrolysis of stachyose to manninotriose seems to occur along the transport path. Based on the specificities of the observed carbohydrate dynamics, the putative physiological roles of manninotriose in red deadnettle are discussed., Conclusions: It is demonstrated for the first time that manninotriose is a novel and important player in the RFO metabolism of red dead deadnettle. It is proposed that manninotriose represents a temporary storage carbohydrate in early-spring deadnettle, at the same time perhaps functioning as a membrane protector and/or as an antioxidant in the vicinity of membranes, as recently suggested for other RFOs and fructans. This novel finding urges further research on this peculiar carbohydrate on a broader array of RFO accumulators.

Tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), overwinter as diapausing adults in North America. Overwintering adults were collected near Stoneville, MS from blooming henbit, Lamium amplexicaule L., and from plant debris during December and January and dissected to determine their reproductive status. Averaged over four winters, male and female tarnished plant bugs collected from henbit terminated diapause at a significantly higher rate than males and females from plant debris during each week of December and the first week of January. Both sexes in each habitat were nearly all reproductive by the end of January. Adults overwintering in plant debris terminated diapause during January in the absence of a food stimulus in all 5 yr studied. This emergence was thought to be controlled by an internal clock. Laboratory and field studies showed that emergence from diapause could be influenced by food, sex, and temperature. Adults overwintering on a suitable food source, blooming henbit, terminated diapause during December in the 4 yr studied, and males terminated diapause more rapidly than females. Food quality was important in emergence from diapause, and females on blooming henbit terminated diapause at a significantly higher rate than females on nonblooming mustard, Brassica juncea (L.) Cosson. Laboratory tests showed that diapausing adults reared in the laboratory and held at a diapause-maintaining photoperiod of 10:14 (L:D) h could be terminated from diapause by using food and temperature stimuli. The lower thermal threshold for development to reproductive adults was found to be near 10°C. The ability of diapausing adults to respond to food and temperature stimuli in December can enable tarnished plant bugs to take advantage of warm winters and winter hosts to produce a new generation earlier.

Koromiko [Hebe salicifolia G. Forst. (Pennell)] is a woody angiosperm native to New Zealand and Chile. Hebe spp. belong to the otherwise herbaceous family Plantaginaceae in the order Lamiales. Reaction wood exerting expansional forces was found on the lower side of leaning H. salicifolia stems. Such reaction wood is atypical for angiosperms, which commonly form contracting reaction wood on the upper side of leaning stems. Reaction wood typical for angiosperms is formed by species in other families in the order Lamiales. This suggests that the form of reaction wood is specific to the family level. Functionally the reaction wood of H. salicifolia is similar to that found in gymnosperms, which both act by pushing. However, their chemical, anatomical and physical characteristics are different. Typical features of reaction wood present in gymnosperms such as high density, thick-walled rounded cells and the presence of (1 → 4)-β-galactan in the secondary cell wall layer are absent in H. salicifolia reaction wood. Reaction wood of H. salicifolia varies from normal wood in having a higher microfibril angle, which is likely to determine the direction of generated maturation stresses.

• Some plants secrete coloured nectar to attract pollinators, but little is known about the chemical origins of nectar colouration and its ecological function. Leucosceptrum canum stands out as the only plant with coloured nectar recorded in the Himalayas. Here, we focused on the compound associated with the dark colour of the nectar, as well as its secretion dynamics during the flowering season and its relationship to pollinators. • Fresh nectar was analysed by semi-preparative reversed-phase high-performance liquid chromatography (HPLC), LC-MS and HRESIMS (high resolution electronspray ionization mass spectroscopy) to determine which compound causes the nectar colouration. Behavioural experiments were conducted with birds and honeybees to elucidate the effect of the nectar colour and volume on pollinators. • We identified a purple anthocyanidin, 5-hydroxyflavylium, as a natural nectar product for the first time. Two short-billed birds were found to pollinate this plant, which employs two nectar-based mechanisms to direct bird pollinators to reproductively active flowers, controlling nectar palatability and presenting a foraging signal for birds by altering nectar volume and colour in a developmental stage-specific manner. • 5-Hydroxyflavylium was found to be the cause of the nectar colouration, the function of which is to act as a foraging signal to increase pollination efficiency through nectar visibility and palatability., (© 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.)

• Understanding how pollination affects plant reproductive success and how changes in pollination service affect plant populations, communities and ecosystems is of increasing concern. Yet supplemental hand-pollination traditionally used to assess pollen limitation is prohibitive for large-scale comparative work. Moreover, it does not differentiate between quality and quantity aspects of pollen limitation, and it may suffer from confounds of post-pollination processes such as resource availability to fill seeds. • Here, we highlight pollen tubes as the functional link between pollen arrival and seed production and suggest that consideration of pollen tubes leads to a better depiction of limitation at the pre-zygotic (pollination) phase of sexual reproduction. • We assessed the rigor of piecewise regression to analyze the relationship between the numbers of pollen grains and pollen tubes observed in nonmanipulated wilted flowers. We illustrate how parameters obtained from this analysis provide quantitative insight into the relative relevance of the quantity and quality of pollen receipt in limiting natural pollination success, and can facilitate comparisons among data sets. • This nonmanipulative method opens up new opportunities for rigorous assessment of the relative importance of the quantity and quality of pollination in limiting plant reproduction, especially from a community-wide perspective., (© 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.)

Hydroponic experiments were conducted to investigate the tolerance and spatial distribution of lead (Pb) in Elsholtzia splendens-a copper (Cu) accumulator plant using synchrotron-based micro-X-ray fluorescence. According to chlorophyll concentration and chlorophyll fluorescence parameters, E. splendens displayed certain tolerance at 100 μM Pb treatment. Lead concentration in roots, stems and leaves of E. splendens reached 45,183.6, 1657.6, and 380.9 mg kg(-1), respectively. Pb was mostly accumulated in the roots, and there were also high concentrations of Pb been transported into stems and leaves. Micro-XRF analysis of the stem and leaf cross section revealed that Pb was mostly restricted in the vascular bundles and epidermis tissues of both stem and leaf of E. splendens. The correlation between distribution of K, Ca, Zn and Pb were analyzed. There were significant positive correlations (P<0.01) among Pb and Ca, K, Zn distribution both in stem and leaf of E. splendens. However, among the three elements, Ca shared the most similar distribution pattern and the highest correlation coefficients with Pb in both stem and leaf cross section of E. splendens. This suggests that Ca may play an important role in Pb accumulation in stem and leaf of E. splendens., (Copyright © 2011 Elsevier B.V. All rights reserved.)