Dioica Plants Research Articles

The impact of mechanical stress on anatomy, morphology, and gene expression in Urtica dioica L.

Main conclusionMechanical stress induces distinct anatomical, molecular, and morphological changes in Urtica dioica, affecting trichome development, gene expression, and leaf morphology under controlled conditionsThe experiments were performed on common nettle, a widely known plant characterized by high variability of leaf morphology and responsiveness to mechanical touch. A specially constructed experimental device was used to study the impact of mechanical stress on Urtica dioica plants under strictly controlled parameters of the mechanical stimulus (touching) and environment in the growth chamber. The general anatomical structure of the plants that were touched was similar to that of control plants, but the shape of the internodes' cross section was different. Stress-treated plants showed a distinct four-ribbed structure. However, as the internodes progressed, the shape gradually approached a rectangular form. The epidermis of control plants included stinging, glandular and simple setulose trichomes, but plants that were touched had no stinging trichomes, and setulose trichomes accumulated more callose. Cell wall lignification occurred in the older internodes of the control plants compared to stress-treated ones. Gene analysis revealed upregulation of the expression of the UdTCH1 gene in touched plants compared to control plants. Conversely, the expression of UdERF4 and UdTCH4 was downregulated in stressed plants. These data indicate that the nettle's response to mechanical stress reaches the level of regulatory networks of gene expression. Image analysis revealed reduced leaf area, increased asymmetry and altered contours in touched leaves, especially in advanced growth stages, compared to control plants. Our results indicate that mechanical stress triggers various anatomical, molecular, and morphological changes in nettle; however, further interdisciplinary research is needed to better understand the underlying physiological mechanisms.

Two novel PCR-based assays for sexing of Silene latifolia and Silene dioica plants

Silene latifolia and S. dioica are model systems in studies of plant reproduction, chromosome evolution and sexual dimorphism, but sexing of plants based on morphology is only possible from flowering stage onwards. Both species show homogametic females (XX) and heterogametic males (XY).•Here we developed two assays (primer pairs ss816 and ss441) for molecular sexing of S. latifolia and S. dioica, targeting length polymorphisms between the X and Y-linked copies of the spermidine synthase gene.The two assays were successful in identifying known (flowering-stage) males and females from UK and Spanish populations, with an error rate of 3.1% (ss816; successful for both species) and 0% (ss441, only successful for S. latifolia). Our assays therefore represent novel tools for rapid, robust and simple determination of the genotypic sex of S. latifolia and S. dioica.

Drug interaction potential of Ankaferd blood stopper® in human hepatocarcinoma cells.

Ankaferd blood stopper® (ABS) is an herbal extract consisting of mixtures of Alpinia officinarum, Gycyrrhiza glabra, Vitis vinifera, Thymus vulgaris, and Urtica dioica plants and has been used in recent years in Turkish medicine as a hemostatic agent. Despite its extensive usage, there is no information available about the drug interaction in HepG2 cells. The current work evaluated the effect of ABS on the expression of CYP1A1-1A2, CYP2E1, and CYP3A4 isozymes that are primarily involved in drug and carcinogen metabolism. We selected HepG2 cells as in vitro cellular models of the human liver. The cells were treated with different concentrations of ABS [0.25%-40% (v/v)]. A crystal violet staining assay was used to determine the cytotoxicity of ABS. We examined drug-metabolizing enzymes, including 7-ethoxyresorufin O-deethylase (CYP1A1), 7-methoxyresorufin O-demethylase (CYP1A2), aniline 4-hydroxylase (CYP2E1), and erythromycin N-demethylase (CYP3A4), in vitro in HepG2 cells. The expression (mRNA, protein) levels of drug-metabolizing enzymes were analyzed by qPCR and Western blotting, respectively. The EC05 and EC10 values for ABS were 0.37% and 0.52% (v/v), respectively. Therefore, 0.37% and 0.52% (v/v) doses were used for the remaining portion of this study. Investigation of the expression and activity levels revealed that CYP1A1-1A2, CYP2E1, and CYP3A4 activities were not affected by ABS significantly, with qPCR and Western blot results corroborating this result. Our study found that the activity, mRNA, and protein expression levels of CYP isozymes did not change with the application of ABS, suggesting that when humans are exposed to ABS, there may not be any risk associated with clinical drug toxicity, cancer formation, and drug metabolism disorders in humans.

Characterization of silver nanoparticles fabricated by green synthesis using Urtica dioica and Lavandula angustifolia and investigation of antimicrobial and antioxidant

Silver nanoparticles (AgNPs) was synthesized from Urtica dioica (UDANP) and Lavandula angustifolia (LAANP) plants. Silver nanoparticles obtained from green synthesis were characterized by UV-Vis, FTIR, SEM, TEM. The DPPH scavenging activities of UDANP were found as 73.58, 84.61, 100, 100, and 100% at 12.5, 25, 50, 100, and 200 mg/L, respectively. On the other hand, DPPH radical scavenging abilities of LAANP were determined as 62.56, 73.07, 78.71, 86.92, and 100% at 12.5, 25, 50, 100, and 200 mg/L, respectively. The ferrous chelating activities of LAANP were found as 7.96, 17.59, 29.45, 41.63, and 59.44% at 12.5, 25, 50, 100, and 200 mg/L, respectively. Besides UDANP was showed 73.58 and 84.61% metal chelating activity at 12.5 and 25 mg/L, respectively. The antimicrobial activity of UDANP and LAANP was performed using broth-dilution method. After the cell viability test of UDANP and LAANP, E. coli cell growth inhibited as 100%.

Differential distribution of resources for females on a dioecious plant affects the small–scale distribution of male of an oligolectic bee

Females of the solitary digger bee Andrena florea Fabricius, 1793 (Hymenoptera, Andrenidae) nest in aggregations and collect pollen almost exclusively on dioecious plants of the genus Bryonia, making this species a good model to study the relationship between nest density, male density, male behaviour and female lecty. At a study site in the valley of the river Serio, in Italy, an aggregation of this bee showed low density of randomly distributed nests and was closely surrounded by B. dioica plants. Female nectar foraging and male feeding and mate–searching activity, confined to the host plants, peaked at similar hours across the day, while female pollen foraging peaked earlier. Males fed on plants of both sexes but seemed to perch waiting for females more frequently on male B. dioica leaves. Individual males more often visited only one of the male plants, for up to four days; here they did not interact aggressively with conspecifics, suggesting scramble competition in resource–based home ranges and not territoriality. These findings are preliminarily in accordance with the predicted resource–based rendez–vous sites at low nest density for oligolectic bees and the predicted occurrence of scramble competition in case of high male density. Additionally, males would maximize their mating opportunity by mainly perching on male plants, the only source of the most limited resource for females (pollen).

Assessment of genetic diversity, population structure and sex identification in dioecious crop, Trichosanthes dioica employing ISSR, SCoT and SRAP markers

Twenty inter-simple sequence repeat (ISSR) and twenty two start codon targeted (SCoT) primers were employed to analyze genetic diversity and population structure among 52 Trichosanthes dioica Roxb. accessions collected from nine different eco-geographical regions of India. ISSR markers proved to be more informative in genetic diversity assessment and produced higher mean number of polymorphic bands (15.25 with 95.96% polymorphism) and polymorphic information content (PIC) value (0.47) compared to SCoT markers (12.55 polymorphic bands with 92.20% polymorphism and PIC: 0.45). Total genetic diversity (Ht) and genetic diversity within populations (Hs) in T. dioica accessions was found to be very high (0.45 and 0.43, respectively). AMOVA analysis also revealed higher genetic variation within populations (81%) than among them (19%). Among different T. dioica populations, very low genetic differentiation (Gst: 0.05) and high gene flow (Nm: 9.32) were observed. T. dioica populations of Bihar state were found to be highly diverse and Kolkata and Cuttack populations were least diverse. T. dioica male plants were more variable than females. UPGMA, Neighbor-Joining and population structure analyses divided T. dioica populations into three main clusters. First cluster comprised of Meerut population, second cluster included of Cuttack and Kolkata populations and populations of Bihar, Delhi and Kanpur occurred in third cluster. Genetic diversity was found to be strongly positively correlated with the latitude and strongly negatively correlated with annual mean rainfall of different T. dioica cultivated regions. For sex identification, one SRAP primer combination, 'Em-6/Me-4' amplified two molecular markers of around 230 and 290 bp specific to male T. dioica plants of Bihar, Kanpur, North Delhi and Meerut populations and were completely absent from female plants.

Sex-related differences in photoinhibition, photo-oxidative stress and photoprotection in stinging nettle (Urtica dioica L.) exposed to drought and nutrient deficiency

Dimorphic plant species can show distinct nutrient needs due to sex-related differences in nutrient allocation to reproductive structures, which can potentially affect their sensitivity to photoinhibition and photo-oxidative stress. Here, we investigated sex-related differences in the extent of photo-oxidative stress in male and female individuals of U. dioica exposed to a combination of severe drought and nutrient starvation. Male and female individuals of U. dioica subject to severe drought stress were exposed to various levels of nutrient availability. First, a set of plants grown under field conditions and exposed to summer drought was used to test the effects of nutrient supply (given as NPK fertilizer). Secondly, the effects of various phosphate concentrations in the nutrient solution were tested in drought-stressed potted plants. The Fv/Fm ratio (maximum efficiency of PSII photochemistry), photoprotection capacity (levels of carotenoids, including the xanthophyll cycle, and vitamins C and E), and the extent of lipid peroxidation (hydroperoxide levels) were measured. Results showed that an application of the NPK fertilizer to the soil had a positive effect on drought-stressed plants, reducing the extent of lipid peroxidation in both males and females. P deficiency led to residual photoinhibition, as indicated by significant reductions in the Fv/Fm ratio, and enhanced lipid peroxidation in females, but not in males. We conclude that (i) increased nutrient availability in the soil can alleviate photo-oxidative stress in drought-stressed U. dioica plants, and (ii) U. dioica plants show sexual secondary dimorphism in terms of photoinhibition and photo-oxidative stress, but this is only apparent when stress infringed on plants is very severe.

A study on possible use of Urtica dioica (common nettle) plant as polonium (210)Po and lead (210)Pb contamination biomonitor in the area of phosphogypsum stockpile.

The aim of this study was to test a possible use of Urtica dioica (common nettle) plant as a biomonitor of polonium 210Po and lead 210Pb contamination near phosphogypsum stacks by determining concentrations of these radionuclides in samples collected from the area of phosphogypsum stockpile in Wiślinka (northern Poland). The 210Po and 210Pb contents in roots depended on their concentrations in soils. Bioconcentration factor values from soil to root of the plant did not depend on 210Po and 210Pb contents in soils that leads to the conclusion that different polonium and lead species have different affinities to U. dioica plants. The main sources of both analyzed radionuclides in green parts of plants are wet and dry air deposition and transportation from soil. The values of 210Po/210Pb activity ratio indicate natural origin of these radioisotopes in analyzed plants. 210Po and 210Pb concentration in U. dioica roots is negatively weakly correlated with distance from phosphogypsum stockpile.

A study on possible use of Urtica dioica (common nettle) plants as uranium (234U, 238U) contamination bioindicator near phosphogypsum stockpile.

The aim of this study was to determine uranium concentrations in common nettle (Urtica dioica) plants and corresponding soils samples which were collected from the area of phosphogypsum stockpile in Wiślinka (northern Poland). The uranium concentrations in roots depended on its concentrations in soils. Calculated BCF and TF values showed that soils characteristics and air deposition affect uranium absorption and that different uranium species have different affinities to U. dioica plants. The values of 234U/238U activity ratio indicate natural origin of these radioisotopes in analyzed plants. Uranium concentration in plants roots is negatively weakly correlated with distance from phosphogypsum stockpile.

English

Several biological properties are attributed to the Pimenta dioica plants that are native to Central America. Given the possible variabilities in both the morpho-anatomical and chemical aspects of the species that grow in different locations, this study aimed to contribute to the pharmacognostic study of P. dioica leaves from Brazil. Therefore, we morpho-anatomically described the plants, performed a phytochemical screening, analyzed the chemical composition of the essential oil and determined the quality parameters of P. dioica leaves that were collected in Goiás State. The leaves are simple and exhibit a camptodromous-brochidodromous venation pattern. The blades exhibit anomocytic stomata on the abaxial side, a dorsiventral mesophyll, secretory cavities on both sides, and a uniseriate epidermis. Trichomes can be observed in the petiole. The tests revealed the presence of phytochemical saponins, tannins and flavonoids in the samples. The major components of the essential oil were eugenol (60.8%) and myrcene (19.3%). The results regarding the moisture content (7.87% w/w), total ash (8.84% w/w), HCl-insoluble ash (1.74% w/w) and total flavonoids (1.88% w/w) can be used as parameters for quality control of botanical material. Additionally, the results of this study indicates strong similarities between the chemical compositions of the leaves of P. dioica plants that were grown in Goiás, Brazil and those from other parts of Brazil and the world.   Key words: Myrtaceae, essential oils, eugenol, morpho-anatomy, Pimenta dioica

Sex-specific interaction between arbuscular mycorrhizal and dark septate fungi in the dioecious plantAntennaria dioica(Asteraceae)

Male and female plants of dioecious species often differ in their resource demands and this has been linked to secondary sexual dimorphism, including sex-specific interactions with other organisms such as herbivores and pollinators. However, little is known about the interaction between dioecious plants and fungal root endophytes. Plants may be simultaneously colonised by arbuscular mycorrhizal (AM) and dark septate (DS) fungi. While it is well established that AM mutualism involves reciprocal transfer of photosynthates and mineral nutrients between roots of host plants and these fungi, the role of DS fungi remains controversial. Here, we report the temporal and spatial variation in AM and DS fungi in female, male and non-reproductive Antennaria dioica plants in three natural populations in Finland during flowering and after seed production. Females had higher colonisation by AM fungi, but lower colonisation by DS fungi than male and non-reproductive plants. The higher AM colonisation was observed during flowering, and this difference varied among populations. Our results suggest that females and males of A.dioica interact with AM and DS fungi differently and that this relationship is dependent on soil fertility.

Differential competitive ability between sexes in the dioecious Antennaria dioica (Asteraceae).

Differences in competitive ability between the sexes of dioecious plants are expected as a result of allocation trade-offs associated with sex-differential reproductive costs. However, the available data on competitive ability in dioecious plants are scarce and contradictory. In this study sexual competition was evaluated using the dioecious plant Antennaria dioica in a common garden transplantation experiment. Male and female plants were grown for 3 years either in isolation, or in competition with a plant of the same sex or the opposite sex. Flowering phenology, sexual and asexual reproduction, plant growth, nutrient content and arbuscular mycorrhizal colonization in the roots were assessed. Our results showed little evidence of sexual differences in competitive ability. Both sexes suffered similarly from competition, and competitive effects were manifested in some traits related to fitness but not in others. Survival was unaffected by competition, but competing plants reduced their vegetative growth and reproductive investment compared with non-competing plants. In addition, differences in sexual competitive ability were observed in relation to flowering frequency, an important life history trait not reported in previous studies. The findings indicate that female and male A. dioica plants possess similar intersexual competitive abilities which may be related to the similar costs of reproduction between sexes in this species. Nevertheless, intrasexual competition is higher in females, giving support for asymmetric niche segregation between the sexes.

Detection and elimination of Begomovirus infecting Trichosanthes dioica (pointed gourd) plants in Uttar Pradesh, India

Begomovirus is widely spread on cultivated crops in India. In a survey, infected plants of Trichosanthes dioica (Pointed gourd) with the symptoms of mosaic and leaf curling were collected and checked for Begomovirus infection through PCR analysis. Application of meristem culture technique for regeneration of virus free plants of T. dioica was assessed. Plantlets were regenerated from infected plants of T. dioica through meristem culture. Regenerated plants were found Begomovirus free as evident by PCR analysis suggesting the suitability of meristem culture technique for elimination of Begomovirus from infected plants of T. dioica. This is the first report on elimination of Begomovirus in T. dioica in India.

Identification of sex in spiny gourd (Momordica dioica) using RAPD marker analysis and its validation in four F1 progenies

SummaryMomordica dioica is an important summer vegetable crop in India and is dioecious. DNA bulks of six male and six female accessions of M. dioica were subjected to random amplified polymorphic DNA (RAPD) marker analysis using 40 primers, of which 22 primers responded for amplification. Of these, ten primers generated reproduction typespecific amplification of DNA fragments at the bulk level. This number was reduced to three RAPD primers when re-tested, individually, with six male and six female accessions. One random decamer primer, OPA-01, produced two unique amplified fragments in all female plants, while two other primers, OPB-04 and OPC-08, generated distinct amplified fragments in both male and female plants. Three amplified fragments (OPB-04430, OPC-081135, and OPC-08578) were unique to all six male accessions, while five amplified fragments (OPA-011560, OPA-01880, OPB-04880, OPB-04800, and OPC-081230) appeared uniquely and uniformly in all six female accessions. Southern hybridisation analysis was performed using five of these sex-specific amplified fragments (OPA-01880, OPB-04880, OPB-04430, OPC-08578, and OPC-081230) as probes with Eco RI and Hind III double-digested DNA of all male and female accessions. Southern hybridisation analysis with the OPC-08578 probe showed a unique hybridisation pattern for all six male accessions, as with the respective RAPD marker, whereas the other probes showed complex hybridisation patterns. Further, inheritance studies also confirmed the presence of a homologous restriction fragment targeted by this probe in F1 progenies, as expected. Hence, OPC-08578 could be used as a marker to ascertain the sex of M. dioica plants at an early stage of development.

Establishment of an efficient Agrobacterium tumefaciens-mediated leaf disc transformation of spine gourd (Momordica dioica Roxb. ex Willd)

Spine gourd ( Momordica dioica Roxb. ex Willd) is a medicinally and economically important plant and also used as vegetable. In this study, we established an Agrobacterium tumefaciens -mediated transformation procedure for M. dioica . Leaf explants were incubated with A. tumefaciens strain LBA4404 containing a binary vector pBAL2 carrying the reporter gene β-glucuronidase intron (GUS-INT) and the marker gene neomycin phosphotransferase (NPTII). Following co-cultivation, leaf explants were cultured on Murashige and Skoog (MS) + Gamborg et al., (B 5 ) medium supplemented with 6.6 μM 2,4- dichlorophenoxyacetic acid (2,4-D) combined with 3.3 μM 6-benzylaminopurine (BAP) containing 100 mg L –1 kanamycin and 300 mg L -1 carbenicillin. Kanamycin-resistant calluses were induced from the leaf explants after three weeks. Shoot regeneration was achieved after transferring the calluses onto fresh selection medium 8.8 μM BAP and 2.2 μM 2,4-D. Transgenic shoots were excised from callus and elongated in MS medium fortified with 3.0 μM gibberellic acid (GA 3 ), 100 mg L -1 kanamycin and 300 mg L -1 carbenicillin. Finally, the shoots were rooted on MS basal medium supplemented with 3.0 μM indole 3-butyric acid (IBA) and 100 mg L -1 kanamycin. High transformation frequency was achieved by using three-day-old precultured leaf explants. Furthermore, the presence of acetosyringone (200 μM), infection of explants for 30 min and three days of cultivation proved to be critical factors for greatly improving the transformation efficiency. Incorporation and expression of the transgenes confirmed by polymerase chain reaction (PCR), Southern blot analysis, reverse transcription (RT)-PCR and GUS histochemical assay. Using this protocol, transgenic M. dioica plants can be obtained in approximately three months with a high transformation frequency of 9%. Key words : Agrobacterium tumefaciens, acetosyringone, growth regulators, GUS, genetic transformation, Momordica dioica .

Sex ratio and spatial distribution of male and female Antennaria dioica (Asteraceae) plants

Sex ratio, sex spatial distribution and sexual dimorphism in reproduction and arbuscular mycorrhizal colonisation were investigated in the dioecious clonal plant Antennaria dioica (Asteraceae). Plants were monitored for five consecutive years in six study plots in Oulanka, northern Finland. Sex ratio, spatial distribution of sexes, flowering frequency, number of floral shoots and the number and weight of inflorescences were recorded. In addition, intensity of mycorrhizal fungi in the roots was assessed. Both sexes flowered each year with a similar frequency, but the overall genet sex ratio was strongly female-biased. The bivariate Ripley’s analysis of the sex distribution showed that within most plots sexes were randomly distributed except for one plot. Sexual dimorphism was expressed as larger floral and inflorescence production and heavier inflorescences in males. In addition, the roots of both sexes were colonised to a similar extent by arbuscular mycorrhizal fungi. The female sex-biased flowering ratios reported are not consistent among years and cannot be explained in terms of spatial segregation of the sexes or sex lability. The possible reasons for the female-biased sex ratio are discussed.

Influence of stress history on the response of the dioecious plant Urtica dioica L. to abiotic stress

Background: Reaching maturity is thought to affect plant responses to stress, but few studies have examined thus far how stress periods during the juvenile phase can alter the response of plants in a mature stage, particularly in dioecious plants. Aims: This study tested the hypothesis that sex and stress history can alter the response of Urtica dioica plants to abiotic stress. Methods: Stress indicators (water content, chlorophylls, the F v/F m ratio and the extent of lipid peroxidation) were measured in leaves of juvenile and mature plants exposed to a combination of water and nutrient deficit, with an emphasis on evaluating the effects of stress during the juvenile phase on the stress response of mature plants. Results: Stress treatment during the juvenile phase affected plant response to stress during the mature phase. Leaves of reproductive shoots were the most sensitive to stress history, as shown by increases in lipid peroxidation in leaves of reproductive shoots relative to non-reproductive ones. Leaves of both shoot types responded similarly in males and females, which showed no significant differences in any of the parameters measured. Conclusions: Stress history appears to determine the response of mature U. dioica plants to abiotic stress, plants with a stress history showing acclimation to subsequent stress, leaves of non-reproductive shoots of both males and females being more stress tolerant and allowing plant survival under severe stress.

Sex‐specific responses to mycorrhiza in a dioecious species

In most studies about dioecious plants, the role of arbuscular mycorrhizae (AM) and the potential sex-specific differences between the plant hosts have been overlooked. Because plant sexes frequently differ in drought tolerance and AM fungal colonization provides higher resistance to drought, we investigated whether the relation of mycorrhizal fungi with either male or female Antennaria dioica plants differs using a factorial experiment. We hypothesized that because AM usually increase growth rate and male plants usually grow larger than females, males should gain more benefit from the mycorrhizal symbiosis in terms of mineral nutrition and water supply. Because of higher demands of carbohydrates (C) in males, we expected males to allocate less C resources to the mycorrhizal fungus so that the associated fungi should benefit less of the association with males. In contrast to our initial hypothesis, the male plants, although faster growing under drought, did not gain more symbiosis-mediated benefits than did the females, and both sexes seemed to provide resources equally to their fungal symbiont. Therefore, we conclude that the two plant sexual morphs provide equal amounts of C to their fungal root symbionts and that they can gain specific benefits from the symbiosis, which, however, depend on soil water availability.

Type 1 ribosome-inactivating proteins from Phytolacca dioica L. leaves: differential seasonal and age expression, and cellular localization

The expression of type 1 ribosome-inactivating proteins (RIPs) in Phytolacca dioica L. leaves was investigated. Fully expanded leaves of young P. dioica plants (up to 3 years old) expressed two novel RIPs, dioicin 1 and dioicin 2. The former was also found in developing leaves from adult P. dioica within about two and a half weeks after leaf development, and the latter continuously synthesized, with no seasonal or ontogenetic constraint. Fully expanded leaves from adult P. dioica expressed four RIPs (PD-Ls1-4) exhibiting seasonal variation. RIPs were localized in the extracellular space, in the vacuole and in the Golgi apparatus of mesophyll cells. Dioicin 1 and dioicin 2 showed rRNA N-beta-glycosidase activity and displayed the following properties, respectively: (1) Mr values of 30,047.00 and 29,910.00, (2) pIs of 8.74 and 9.37, and (3) IC(50) values of 19.74 (0.658 nM) and 6.85 ng/mL (0.229 nM). Furthermore, they showed adenine polynucleotide glycosylase activity and nicked pBR322 dsDNA. The amino acid sequence of dioicin 2 had 266 amino acid residues, and the highest percentage identity (81.6%) and similarity (84.6%) with PAP-II from Phytolacca americana, while its identity with other RIPs from Phytolaccaceae was around 40%.

Inheritance of progeny sex ratio inUrtica dioica

Seed samples collected from female Urtica dioica plants in the field showed considerable inter-family variation in the sex ratio (faction of males). To investigate the inheritance pattern of the sex ratio trait, crosses were performed between individual male and female plants from different sex ratio families. Our results suggest, at least for the families studied here, that maternal parents strongly contribute to the variation in the primary sex ratio. Furthermore, progeny sex ratios from reciprocal crosses were significantly different and resembled the sex ratios produced by their maternal parents. We discuss the possible mechanisms underlying maternal control.