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A method of in vitro clonal propagation of cowslip Primula veris L. (Primulaceae) is reported. This is a species of medicinal importance, protected by law in Poland. MS medium Murashige and Skoog (1962) with 6 benzyladenine BA (4.44 µM) and 2,4-dichlorophenoxyacetic acid 2,4-D (1.13 µM) was found to be optimal for in vitro cowslip propagation from shoot tips under the conditions of culture. Rooting was best in the presence of indole-3-butyric acid IBA (2.45 µM). In vitro seedlings were transferred to pots and then acclimated. After transplanting to a garden they showed further growth and development, including flowering and fruiting. Phytochemical analysis (2D-TLC) revealed that the flavonoid compounds in leafy shoots from in vitro culture were similar to those in leaves from field cultivation.
Morphological characterisation and the evaluation of growth and development of cultivated Primula veris L. plants were done according to different origin of seeds which the cultivated populations were started from and to two types of row spacing applied in the experimental plots. For each plant the number of leaves per plant and the rosette diameter were determined. At the flowering phase the number of scapes per plant, as well as the numbers of buds, flowers and fruits per plant and per scape were determined. The level of saponin content, converted to primulic acid, in rhizomes and roots of cowslip plants from natural and cultivated populations was assessed by means of the densytometric method. The growth and development of cowslip plants under cultivation conditions were more intensive at the plant spacing of 40 x 30, whereas fewer leaves dried on plants at the spacing of 30 x 15. The number of leaves and the diameter of leaf rosettes of the studied plants had a significant influence on the number of scapes per plant as well as on the number of flowers and fruits per plant. The saponin content in the underground organs of cowslip plants increased by 0.8-4.0% after transferring plants from natural habitats to cultivation conditions.
Micromorphological characterisation and the comparative statistical analysis of the size of Primula veris L. pollen grains collected in three natural and three cultivated populations were done. Observations were carried out with SEM. The obtained measurements were analysed with the use of one-way ANOVA, Kruskal-Wallis Test and the Student-t Test. Pollen grains from long-styled ('pin') flower-morphs were mainly 6 colpate and from shortstyled ('thrum') flower-morphs 8 colpate. Colpi of some grains from 'thrum' flowers were 'sinuous' and 'circular', and they incised into the apocolpium zone. Ornamentation of 'pin' pollen grains was microreticulate, with lumina up to 0.8 μm wide, and for pollen grains from 'thrum' flowers was reticulate and eureticulate with lumina 1.1-1.7 μm wide. In lumina of mesocolpium area some free columellae were observed. Pollen grains from 'thrum' flower-morphs were more variable in size, both in natural and in cultivated populations, than grains from 'pin' flower-morphs. The differences in mean length (P) and breadth (E) of pollen grains from 'thrum' flowers collected in cultivated populations were statistically important (FP = 3.154 for the critical F005 = 3.098; K-We = 7.469 for the critical Test value α=005 = 5.991). Pollen grains from 'thrum' flowers were bigger when coming from plants growing in natural populations (tE = 2.784 for the critical Test value α=005 = 2.001).
The number and the weight of seeds per capsule were assessed along with the individual seed mass in cowslip plants of various origin, grown in plots with differing plant spacing. The potential seed productivity (the number of ovules per ovary) and the real seed yield (the number of seeds per capsule) were determined for the cultivated plants and the effect of seed origin and gibberellic acid (GAj) on germination capacity was evaluated under field conditions. The mean seed number per cap­sule for plants growing at the 40 x 30 spacing ranged from 31.2 to 64.4, whereas for the 30x15 spacing it ranged from 37.2 to 60.6 seeds per capsule, depending on the year of study and the flo­wer morph type. The seed number per capsule and the individual seed mass turned out to be traits dependent in a statistically significant way on the origin of seeds used to start the cultivated po­pulations. Plants from populations 2 and 3 produced the heaviest seeds (mean = 1.10 mg and 1.14 mg). The differentiation in the number of seeds per capsule and in the individual seed mass in fruits coming from the same scape was statistically significant. The mean number of ovules per ovary ranged from 64.8 to 71.0 and the mean seed number per capsule ranged from 46.2 to 51.7. The number and percentage of germinated .seeds ranged from 6 (4.0%) to 48 (32.0%), depending on the seed origin. Seeds treated with GA3 germinated in 36.0 to 813%.
The reproduction ability of Primula veris L. plants was described on the basis of six-year observa­tions on permanent plots in populations from natural localities. Cowslip individuals in different life stages were mapped according to topographical distribution. The vegetative multiplication symptoms of the plants were also recorded. The mean survival rate of seedlings to the juvenile sta­ge was similar in both populations, amounting to about 16.5%. The highest survival rate was found for juveniles (44.3-57.6%) and immature (31.4-48.6%) individuals, while the lowest number of indi­viduals turned from the virginile into the generative stage (2.2-5.7%). The efficiency of the gene­rative reproduction measured by the ratio of the number of generatively recruited flowering plants to the observed number of seedlings was 0.22-0.42%, depending on the population.
Primula veris L. (Primulaceae) is a well-known medicinal herb. The callus induction response of three explant types: roots, cotyledons, and hypocotyls of four-week-old cowslip seedlings were evaluated. The highest statistically different callus induction rate was 93.6% and was obtained from root explants on MS medium supplemented with 0.1 mg/l BA and 5.0 mg/l PIC. Calli also appeared on 83.3% of cotyledon explants on MS medium supplemented with 1.0 mg/l BA and 3.5 mg/l 2,4-D and on 81.0% of root explants on MS medium supplemented with 0.1 mg/l KIN and 2.0 mg/l 2,4-D. These values were not statistically different. The average time required for callus initiation was 4 to 6 weeks, however, it depended on the explants type. The most suitable condition for callus proliferation and growth was MS medium with 0.5 mg/l TDZ and 0.1 mg/l NAA, and with 1.0 mg/l BA and 2.0 mg/l or 3.5 mg/l 2,4-D. No light conditions proved to be more favourable for cowslip calli induction and growth.
Ascorbic acid is a well-known antioxidant found in plants. The content of ascorbic acid was assayed using a normal phase European Pharmacopoeia HPLC method for ascorbic acid in medicinal products. The content of ascorbic acid in herbs was calculated in % for absolutely dry drug. Ascorbic acid was not detected in the roots of Primula veris, in aerial parts it was detected in flowers (0.43 ± 0.034%), in blades (1.43 ± 0.11%) and petioles (1.56 ± 0.12%). In fresh leaves collected at weekly intervals the content of ascorbic acid varied from 1.19 to 2.39%, being highest from mid-May to mid-June. The fresh leaves contained 2.35 ± 0.18% of ascorbic acid and when frozen its content was quite stable for one year. The content of ascorbic acid in dried leaves decreased more than ten times in three months, in twelve months it was less than 1/20th of the initial level. Compared to the analyzed common fresh fruits and salads (n = 10) the fresh leaves of common cowslip contained considerably more ascorbic acid. Commercial orange juices could be recommended as the most convenient source of ascorbic acid (8.6–50.4 mg/100 ml); 1–5 glasses of orange juice could fulfill the recommended daily intake of vitamin C (60 mg).
Genetic variation of twelve Polish populations of Primula veris L. from western Poland was investigated in respect of six enzyme systems: 6-phosphogluconate dehydrogenase (6PGD), diaphorase (DIA), menadione reductase (MNR), formate dehydrogenase (FDH), isocitrate dehydrogenase (IDH) and glutamate oxaloacetate transaminase (GOT). Only two of them (6PGD and DIA) were polymorphic and all populations were compared according to four loci and eight alleles. For 6PGD only one out of the two detected loci (locus 6PGD-2) was polymorphic and consisted of three alleles a, b and c. For DIA each of two detected loci had two alleles. For 6PGD-2 one population was monomorphic and four populations were monomorphic for DIA-1 and DIA-2. The rest of the populations were polymorphic with low frequency of heterozygotes. The low heterozygosity level, found in the examined populations, was confirmed by high values of the fixation index (F). The level of genetic differentiation among GST populations specified for each polymorphic loci, was equal to 0.045 for 6PGD-2 and had the value of 0.078 for DIA-2 and 0.186 for DIA-1. Nm value for polymorphic loci was 1.10 for DIA-1 and 2.94 for DIA-2, and for 6PGD-2 was 5.33, what indicates some gene flow between the examined populations. The dendrogram constructed on the basis of genotype frequencies showed that the populations were divided into two groups, however the most southern population No. 2 was clearly similar to the northern population No. 8.
Primula veris, a well-known pharmaceutical plant, is a long-lived perennial protected by law in Poland, so its rhizomes and roots can be collected as pharmaceutical stock only from cultivation. Genetic variation of three natural P. veris populations numbering 500-1200 individuals and of three cultivated populations derived from seeds collected from natural sites was investigated in respect of two enzyme systems: phosphogluconate dehydrogenase (6PGD) and diaphorase (DIA). Four presumptive loci were identified from these two enzyme systems. In 6PGD, only one (6PGD-2) of two detected loci was polymorphic, consisting of three alleles a, b and c. Each of two electrophoretically detected loci in DIA was polymorphic and had two alleles. Comparison of the cultivated and natural populations revealed slight differentiation in the presence and composition of genotypes for 6PGD-2, while for DIA all populations except one preserved the same set of genotypes. Mean values of the polymorphism index for three loci ranged from 0.239 to 0.345 for natural populations and from 0.303 to 0.446 for cultivated populations, indicating that cultivated populations were more polymorphic than natural ones. The level of heterozygosity in the examined populations was very low. Mean values for Ho calculated for three polymorphic loci ranged from 0.033 to 0.056. The observed low heterozygosity level was confirmed by high values of Wright’s fixation index, ranging from 0.798 to 0.910.
Zasadność stosowania leków roślinnych w chorobach układu oddechowego wynika z tradycji i w wielu przypadkach dobrze udokumentowanego działania. W niniejszej pracy zaprezentowano ich podział na leki hamujące odruch kaszlowy, działające do nich przeciwnie, tzn. wykrztuśnie, przedstawiono także surowce o silnym działaniu przeciwbakteryjnym i takie, które powodują efekt spazmolityczny w mięśniach gładkich dróg oddechowych.
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