Annals of West University of Timişoara, ser. Biology, 2016, vol. 19 (1), pp.77-86 BELLIS PERENNIS - VARIATIONS OF PHYSIOLOGICAL RESPONSES IN URBAN CONDITIONS Lorena Alina CIOBANU Department of Biology and Chemistry; West University of Timisoara, Romania Corresponding author e-mail: [email protected] Received 26 February 2016; accepted 14 April 2016 ABSTRACT In the present study, Bellis perennis were sampled from park and a nearby street in the urban area of Timisoara and in different time periods, i.e. April and July. The objectives were to examine the potential impacts of traffic pollution on the B. perennis and the repartition of water, dry matter and organic matter into different plant parts (leaves, scapes, inflorescences and roots). Our results would be helpful in understanding the resource distribution within the plant in urban environment. KEY WORDS: physiological parameters, traffic site, Bellis perennis INTRODUCTION Bellis perennis L. (common daisy) is a perennial herbaceous plant and member of the Asteraceae family. It is widely distributed in Europe and North Africa, but is commonly found as an invasive plant in North America, South America and New Zealand (Tutin et al. 1976; Brouillet, 2006; Zangenehgheshlaghi et al . 2012; Smith et al. 2013; Al-Snafi, 2015). This herbal medicine is also well-known as an ornamental plant (Ramezanzadeh et al. 2014; Ianovici, 2015). Selected strains are cultivated for decoration (Davis, 1995). Its flowers and young leaves are edible as a salad (Koca et al. 2015). The main constituents of this plant, several triterpenoid saponins (Hiller et al. 1988; Schöpke et al. 1991), anthocyanins (Toki et al. 1991), flavonoids (Gudej & Nazaruk, 2001), polyacetylenes (Avato et al. 1997), tannins (Hegi, 1979), essential oil (Tava, 1996) and phenolic acids (e.g., caffeic, ferulic, sinapic, p-coumaric, and salicylic acids) (Grabias et al. 1995) have been isolated from the roots and flowers. The whole flowering plant of B. perennis has been used for bruises, bleeding, muscular pain, purulent skin diseases, and rheumatism in traditional medicine (Morikawa et al. 2011). B. perennis has been used as a diuretic, antispasmodic, anti-inflammatory, astringent, expectorant, antipyretic, vulnerary, ophthalmic and homeostatic (Grieve, 1982; Bown,1995; Baytop, 1999; Duke et al. 2002). The methanolic extract and its saponin constituents were found to show inhibitory effects on plasma triglyceride elevation in olive oil-loaded mice (Morikawa et al. 2008) and pancreatic lipase inhibitory activity (Morikawa et al. 2009). Antibacterial, antifungal, antioxidant (Desevedavy et al. 1989; Kavalcioglu et al . 2010a), postpartum antihemorrhagic (Oberbaum et al. 2005), and cytotoxic activities against HL-60 human promyelocytic leukemia cells (Li et al. 2005) of B. perennis have also been shown. The methanol extract of B. perennis flowers has anti- proliferative effect both on human breast cancer (MCF-7) and human hepatocellular carcinoma (HepG2/C3A) cancer cells (Karakaș et al . 2015). B. perennis may produce biphasic or bipolar effects on learning performance (Karakaș et al. 2011). Traditional usage of wound healing activity of B. perennis was scientifically verified (Karakaș et al . 2012). 77 CIOBANU: Bellis perennis - variations of physiological responses in urban conditions Physiological and growth responses of B. perennis to temperature (Gunn & Farrar, 1999), UV-A and UV-B irradiances (Cooley et al . 2000a, Cooley et al. 2000b) have been described. Ultraviolet light influences dry weight of flowers, leaves, and roots, leaf area, photosynthetic parameters, and transpiration rate depending on wavelength and intensity of radiation, and temperature affects plant total dry mass, leaf area, and root respiration. The flowering heads are nyctinastic, closing from dusk to possibly mid-morning on a daily basis (Faur & Ianovici, 2004). Scognamiglio et al. (2012) study evidenced a potential allelopathic role of caffeic acid derivatives from B. perennis based on inhibitory or stimulatory effects on wild coexisting species growth. Contents of phenolics and flavonoids as well as radical scavenging activity of daisy flowers vary to a relatively small extent during the year and are not dependant on the time of collection (Siatka & Kašparová, 2010). On the basis of Kavalcioglu et al . (2010b) results it is clear that there are significant variations in the field collected populations of B. perennis from different geographic and climatic locations. Street plants are exposed to a relatively high stress level, including high pollutant concentrations (Jim, 1998; Ianovici, 2007; Berlizov et al, 2007; Ianovici et al. 2009; Day et al. 2010; Harris & Manning, 2010; Ianovici et al. 2012; Demuzere et al. 2014; Ianovici et al. 2015a; Calfapietra et al. 2015; Ianovici & Latiș, 2016). The goal of this work was to analyze the physiological parameters of B. perennis during the flowering stage and possible adaptations (Ianovici et al. 2011; Ianovici et al. 2015b) to environmental conditions. MATERIALS AND METHODS In the present study, B. perennis were sampled from park (background site -BS) and a nearby street (traffic site - TS) in the urban area of Timisoara and in different time periods, i.e. April and July. All plants of B. perennis were cut during spring (late April) and summer (mid- July). Plant materials were collected in flowering stage. Rosette populations grow logistically, probably regulated by change in birth rate. The plants remain winter green and continue to grow (Schmid & Harper, 1985; Grime et al. 1988). The species has a very long flowering season, with flowers being produced mainly from about March to November (Mitich, 1997). B. perennis is often common in grasslands and they are generally in shady habitats. This plant prefers a pH of 7.0 to 8.0. At each harvest, plants were separated into roots, scapes, leaves and inflorescences. All samples were individually weighed fresh in the laboratory (FW in g), then oven-dried and re-weighed to estimate their biomasses (DW in g). All plant parts were incinerated at 500°C for 2 hours before recording the ash weights at analytical balance (AC in g). Organic matter (OC in g) is left when the ash content subtract from dry matter (Grudnicki & Ianovici, 2014). List of non-standard abbreviations of physiological parameters calculated: IWC (%) - initial water content; MC (%) - mineral content; OC (%) - organic content; OC/MC ratio - organic content/ mineral content ratio; LRWC (%) - leaf relative water content; TD (g/kg) - tissues density; TDM (g/kg dry wt) - tissues mineral deposition. At the level of leaves we determined the other two parameters: leaf relative water content (LRWC in %) and specific leaf area (SLA in cm -2/g -1) (Cornelissen et al. 2003; Ianovici, 2011a; Ianovici, 2011b; Ianovici, 2016). Statistical analyses were performed with the software SPSS. Probability ( p) values less than 0.05 were considered significant. The data had a normal distribution. Variance between atmospheric fungal spores distributions was analyzed by one-way ANOVA between groups, 78 Annals of West University of Timişoara, ser. Biology, 2016, vol. 19 (1), pp.77-86 Levene 's test for homogeneity of variance (based on means) and Welch F test in the case of unequal variances. Analysis of significant differences between AFS was followed by Tukey HSD (honest significant difference) post-hoc test (Oehlert, 2010; Ianovici et al. 2013). RESULTS AND DISCUSSIONS The analyzed physiological parameters of leaves are reported in Table 1. Regarding the leaves, the data were homogeneous for the following parameters: FW, DW, AC, OC, SLA, TD, LOC/LMC ratio, IWC and OC in %. Analysis of variance revealed that the data are significantly different for SLA, LOC/LMC ratio and OC in % fresh wt. When the data were heterogeneous, we applied Welch F test. In this case, the F was significantly different for TMD and MC in % fresh wt. Fresh leaf weight ranged between 0.0611g and 0.1120g, and dry weight between 0.0065g and 0.0111g. Organic content of samples in the summer increased compared with the spring samples. Both LRWC and IWC values were slightly lower in summer. Tukey’s pairwise comparisons for LOC/LMC ratio indicated significant differences for the following samples: spring/BS – summer/BS, spring/TS – summer/BS, summer/BS – summer/TS. The study of variations of SLA indicated significant differences but Tukey’s pairwise comparisons indicated no significant differences for the samples. However, the SLA values are higher in summer (489.6525 cm -2/g -1in background site and 441.4667 cm -2/g -1in traffic site). The results for TMD and MC fell in summer. TMD and MC values in traffic sites are greater than the background sites, in both seasons. For scapes, data were homogeneous for the following parameters: FW, TD, LOC/LMC ratio, IWC, MC in % fresh wt and OC in % (table 2). Analysis of variance revealed that the data are significantly different for FW, LOC/LMC ratio, MC in % fresh wt and OC in % fresh wt. Welch T test did not show significantly different values for the other parameters. Fresh and dry weight of scapes in summer was lower than in spring. Tukey’s pairwise comparisons for FW indicated significant differences for the following samples: spring/BS – spring/TS (p=0.02817), spring/BS - summer/BS (p=0.001224), spring/BS – summer/TS (p=0.00365). Between samples from background sites we identified significant differences for TD (p=0.03699), IWC (p=0.03699) and OC% (p=0.03861). Results for the TD and OC in % were higher in summer. IWC values fell in the summer. TMD and MC values in summer/TS are significantly higher than all other types of samples. Levene's test indicated that all parameters for inflorescences were homogeneous (table 3). Analysis of variance revealed that the data are significantly different for FW, AC and OC / MC ratio. The inflorescences were heavier the spring / BS (0.157933g). The Tukey's pairwise comparisons for FW indicated significant differences for samples: spring / BS - spring / TS (p=0.0179), spring / BS - summer / BS (p=0.01885), spring / BS - summer / TS (p=0.02215).