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Antimitotic and antibacterial effects of the Primula veris L. flower extracts

Authors:
Gamze Başbülbül at Aydın Adnan Menderes University
Gamze Başbülbül
Ali Ozmen at Aydın Adnan Menderes University
Ali Ozmen
Halil Biyik at Aydın Adnan Menderes University
Halil Biyik
Özge Şen
Özge Şen
Özge Şen
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Primula is a plant genus which comprises about 400 species. It has been found in a number of pharmacological studies that primrose extracts are rich in saponins. Phenolic glycosides and saponins are characteristic compounds for the genus Primula. In this study several flower extracts from Primula veris L. has been tested for antibacterial activity and decoction from the flowers has been tested for antimitotic activity. Antibacterial activity was determined by the well diffusion method and Allium cepa L. has been used for evaluating cytotoxicity. Decoction of flowers was toxic on root number and root length in A. cepa L. and reduced the mitotic index significantly. All of the tested P. veris L. extracts showed inhibitory effect against both Gram positive and Gram negative microorganisms at varying degrees. The most effective fraction was found to be the ethanolic.
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Caryologia
International Journal of Cytology, Cytosystematics and Cytogenetics
ISSN: 0008-7114 (Print) 2165-5391 (Online) Journal homepage: http://www.tandfonline.com/loi/tcar20
Antimitotic and antibacterial effects of the Primula
veris L. flower extracts
Gamze Başbülbül , Ali Özmen , H. Halil Biyik & Özge Şen
To cite this article: Gamze Başbülbül , Ali Özmen , H. Halil Biyik & Özge Şen (2008) Antimitotic
and antibacterial effects of the Primula veris L. flower extracts, Caryologia, 61:1, 88-91, DOI:
10.1080/00087114.2008.10589614
To link to this article: http://dx.doi.org/10.1080/00087114.2008.10589614
Published online: 04 Feb 2014.
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Citing articles: 2 View citing articles
CARYOLOGIA Vol. 61, no. 1: 88-91, 2008
INTRODUCTION
Primula is a plant genus included about 400
species. Some of them are popular garden plants
because of their colourful blossoms. Efficacy of
primrose extracts which are rich in saponins have
been demonstrated in a number of pharmaco-
logical studies, which has potent anti-asthmatic,
anti-inflammatory and anti-viral properties. Phe-
nolic glycosides and saponins are characteristic
compounds for the genus Primula (MÜLLER et al.
2005). Flavonoids may have existed in nature for
over one billion years. Methoxyflavones have im-
portant effects in plant biochemistry and physi-
ology, acting as antioxidants, enzyme inhibitors,
precursors of toxic substances and have long been
recognized to possess anti-allergic, anti-inflamma-
tory, antiviral, anti-proliferative and anti-carcino-
genic activities as well as to affect some aspects of
mammalian metabolism (HUCK et al. 2000). Ten li-
pophilic flavones were isolated from Primula veris
L. in vitro cultures (BUDZIANOWSKI et al. 2005).
Two new flavonol glycosides have been identified
and isolated from Italian Primula species (FICO et
al. 2007). Otherwise Primula veris L. has a poten-
tial anxiolytic activity (SUFKA et al. 2000). Primula
species can also contains allergens (PAULSEN et al.
2006) and some species are used traditionally to
treat epilepsy and convulsions (JAGER et al. 2006).
Another Primula species has flavonoids that pos-
sessed strong cytostatic properties against HL 60
cells even at low concentrations (TOKALOV et al.
2004). The biological effects of the genus Primula
are evident. The aim of this study is to determine
antimitotic and antibacterial effects of several
flower extracts from Primula veris L..
MATERIAL AND METHOD
Extraction of flowers - Ether extract: 20 g of dried
and milled flower were placed in a soxhlet car-
tridge and extracted with diethylether at 350C.
After extraction Ether was evaporated by a rotary
evaporator connected to a vacuum pump.
Ethanol extract: The residue in soxhlet car-
tridge has been dried and treated with ethanol in
a shaker at room temperature. After extraction
ethanol was evaporated.
Decoction - 50 g of dried and milled flower has
been boiled in 1000 ml distilled water for 1 h.
After boiling the extract was filtered and a part
of this filtrate has been freeze-dried for preparing
the Water extract. After freezing the water was
removed by lyophilization.
Antimitotic and antibacterial effects of the Primula veris L.
flower extracts
Gamze Bas¸bülbül, Ali Özmen*, H. Halil Biyik and Özge S¸en
Adnan Menderes Üniversitesi, Fen-Edebiyat Fakültesi Biyoloji Bölümü Aydýn, Turkey.
Abstract — Primula is a plant genus which comprises about 400 species. It has been found in a number of phar-
macological studies that primrose extracts are rich in saponins. Phenolic glycosides and saponins are characteristic
compounds for the genus Primula. In this study several flower extracts from Primula veris L. has been tested for
antibacterial activity and decoction from the flowers has been tested for antimitotic activity. Antibacterial activity
was determined by the well diffusion method and Allium cepa L. has been used for evaluating cytotoxicity. Decoc-
tion of flowers was toxic on root number and root length in A. cepa L. and reduced the mitotic index significantly.
All of the tested P. veris L. extracts showed inhibitory effect against both Gram positive and Gram negative micro-
organisms at varying degrees. The most effective fraction was found to be the ethanolic.
Key words: Allium cepa L., antibacterial, antimitotic, cytotoxicity, Primula veris L..
* Corresponding author: phone: ++90 256 2128498;
fax: ++90 256 2135379; e-mail: aozmen@adu.edu.tr
Downloaded by [181.112.228.71] at 19:15 28 January 2016
antimitotic and antibacterial effects of the
primula veris
l. flower extracts 89
Antimitotic activity - Allium cepa has been used
for evaluating cytotoxic properties since the ear-
ly 1920’s (GRANT 1982). This method is an easy
and sensitive tool for measuring the total toxic-
ity caused by chemical treatments as expressed
by growth inhibition of the roots of onion bulbs.
It has been reported that the results from Al-
lium test fit in well in a test battery composed of
prokaryotes and /or other eukaryotes (FISKESJÖ
1993). Small onion bulbs are carefully unscaled
and cultivated on top of test tubes filled with the
decoction of flowers. Water was used as a con-
trol. The test tubes were kept in an incubator at
24±20C and the test samples were changed dai-
ly. After 72 h the roots were counted and their
lengths were measured for each onion. When the
newly emerged roots measured 2.0 – 3.0 cm, they
were fixed. The fixative solution was glacial acetic
acid/absolute alcohol (1/3 v/v). The root tips were
kept in aceto-alcohol solution for 24 h. After fixa-
tion, the slides were prepared for examination or
the roots were transferred to %70 ethyl alcohols
and stored in a refrigerator. For examination, the
root tips were put into a watch glass to which 9
drops of aceto-orcein and 1 drop of 1 M HCl were
added and warmed over a flame of spirit lamp for
2-3 min. These tips were kept at room tempera-
ture for 15-30 min. After removing the root caps
from well-stained root tips, 1 mm of the mitotic
zones were immersed in a drop of %45 acetic-
acid on a clean slide and squashed under a cover
glass. In order to spread the cells evenly on the
surface of the slide, squashing was accomplished
with a bouncing action by striking the cover glass
with a match stick. MI was expressed in terms of
divided cells/total cells. A statistical analysis was
performed on the collected data. The means of
the control and seed extracts were obtained from
descriptive analysis and an Independet-samples
test was performed to obtain P values.
Antibacterial activity - Antibacterial activity was
determined by the well diffusion method. Muller
Hinton agar plates were seeded with 24 h cul-
tures of the bacterial strains. The inoculum was
adjusted to 0.5 MacFarland turbidity standards
(108 cfu/ml). Muller-Hinton Agar plates were in-
oculated with each of these bacterial suspensions
using sterile swabs. The dried plant extracts were
dissolved in sterile dimethylsulfoxide (DMSO)
to give a final concentration of 100 mg/ml. Wells
were cut into the agar and filled with 50 ml of the
plant extracts. Sterile DMSO was used as nega-
tive control. Inoculated plates were incubated at
37°C for Staphylococcus aureus ATCC 25923, Es-
cherichia coli ATCC 35218, Enterococcus faecalis
ATCC 51299, Proteus sp., Listeria sp. , Serratia
marcescens and at 30 °C for Micrococcus luteus
ATCC 9341, Bacillus cereus ATCC 11778, Pseu-
domonas fluorescens DSMZ 50090, B. sphaericus
DSMZ 396.
The antibacterial activity was evaluated by
measuring the diameter of inhibition zone. The
experiment was carried out in duplicate and the
mean of the diameter of the inhibition zones was
calculated.
RESULTS AND DISCUSSION
Antimitotic activity - The root lengths and num-
bers from control and decoction are given in ta-
ble 1. Primula veris L. flower decoction reduced
significantly root number and root length when
compared with control.
Table 1 — The average root lengths and numbers in
control and in decoction after 72 h.
Extract Average root
numbers
(±SD)
Average root
lengths (mm)
(±SD)
Control 35 (±4) 27.9 (±3.7)
Decoction 27.8 (±3.8)* 7.1 (±1.7)*
*Significant at 0.05 level
These results show that the extract from
Primula veris L. flowers has inhibitory effects on
root growth and length in Allium cepa. In con-
formity with human cell cytotoxicity (TOKALOV et
al. 2004) it was found that Primula veris L. flower
decoction has cytotoxic properties also in plant
test systems.
Table 2 — The dividing and total cells that counted
in microscopic observations and mitotic index (MI) in
control and in decoction.
Extract Total cells Dividing cells MI (±SD)
Control 10000 1617 %16 (± 1, 6)
Decoction 10000 516 %5 (± 1, 2)*
*Significant at 0.05 level
In table 2 the mitotic indexes are given for
control and for decoction. It is evident that de-
coction of flowers reduced the mitotic index sig-
nificantly. In conclusion antimitotic effect of plant
is provided by substances which found in flower
decoction.
In respect of this results, Primula veris L. flowers
contains antimitotic constituents that can stop the
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gamze, özmen, biyik and s¸en
90
mitosis in anywhere of the cell cycle. Furthermore
these constituents probably affect the cytoskeleton
or tubulin polymerization or degradation.
Antibacterial activity:
Antibacterial activity of three different ex-
tracts of Primula veris L. has been evaluated in
vitro against ten bacterial test species, which are
known to cause some infections in humans. These
results are given in table 3.
All the tested extracts have inhibited both
Gram positive and Gram negative bacterial species
at varying degrees. Among the tested microorgan-
isms E. faecalis, B. cereus and Pseudomonas fluores-
cens were inhibited by all extracts. Ether and water
extracts have higher inhibitory spectrum from that
of ethanol extract. None of the tested extracts did
show inhibitory effect against S. aureus, Proteus sp.
and Listeria sp. The biggest inhibition zone was
observed with ethanol fraction.
It is known that Primula herb has antispas-
modic, vermifuge, emetic and astringent effect in
public medicine. However, there has been rela-
tively few study in literature about antimicrobial
and anticancer effects of this plant. Primin (2-
methoxy-6-n-pentyl-1, 4-benzoquinone), a natu-
rally occurring product obtained from Primula
obconica has shown antimicrobial and antitumour
properties (BRONDANÝ et al. 2007). An other liter-
ature reports that water insoluble crude extracts
from Primula longipes aerial parts has strong an-
timicrobial activity with low MIC values against
both Gram positive and Gram negative bacteria
(BURUK et al. 2006).
Antimycobacterial effect of Primula has also
been investigated. Leaves and flower extracts of
Primula vulgaris Huds. subsp. sibthorpii has shown
to be active against Mycobacterium tuberculosis
H37RV (ATCC 27294) and extracts caused % 41 in-
hibition of M. tuberculosis (TOSUN et al. 2005).
Traditionally, Primula extracts are prepared
with water in folk medicine and especially con-
sumed as Primula tea. Because of water extract
found as potentially active fraction against many
bacteria, results of this study support the tradi-
tional use of this herb. Additionally, there must be
very active compounds in the other extracts while
they show wide inhibitory spectrum.
The results of these antimicrobial screening
confirms the potential of Primula herb for produc-
tion of bioactive compounds. These findings are
useful tools for rationalizing the use of medicinal
plants in folk therapy. However, the phytochemi-
cal characterization of extracts and identification
of biologically active compounds are necessary.
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BRONDANI D.J., NASCIMENTO C.R.M, MORERIA M.,
LIMA LEITE A.C., DE SOUZA I.A., BIEBER L.W., 2007
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logues. Medicinal Chemistry, 3: 369-372.
BUDZÝANOWSKI J., MOROZOWSKA M. and WESOŁOWSKA
M., 2005 — Lipophilic flavones of Primula veris L.
from field cultivation and in vitro cultures. Phyto-
chemistry, 66: 1033-1039.
BURUK K., SÖKMEN A., AYDIN F., ERTÜRK M., 2006 —
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erapia, 77: 388-391.
FÝCO G., RODONDÝ G., FLAMÝNÝ G., PASSARELLA D.
and TOME F., 2007 — Comparative phytochemical
and morphological analyses of three Italian Primula
species. Phytochemistry, 68: 1683-1691.
FISKESJÖ G., 1993 — Allium Test 1:A 2-3 day plant test
for toxicity assesment by measuring the mean root
growth of onions (Allium cepa L.). Environmental
Toxicology and Water Quality, 8: 461-470.
GRANT W.F., 1982 — Chromosome aberration assays in
Allium. Mutation Research, 99: 273-291.
Table 3 — Diameters of inhibition zones.
Extracts
Zone of inhibition (mm)
S.aureus
E.faecalis
B.cereus
B.sphaericus
M.luteus
S.marcescens
E.coli
Proteus sp.
Listeria sp.
P.fluorescens
Ether – 10 12 11 – – 8 – – 12
Ethanol – 12 8 – – – – – – 28
Water – 12 10 8 – 8 – – – 20
Downloaded by [181.112.228.71] at 19:15 28 January 2016
antimitotic and antibacterial effects of the
primula veris
l. flower extracts 91
HUCK C.W., HUBER C.G., ONGANÝA K.H., Bonn G.K.,
2000 — Isolation and characterization of methoxy-
lated flavones in the flowers of Primula veris by liq-
uid chromatography and mass spectrometry. Journal
of Chromatography A, 870: 453-462.
JAGER A.K., GAUGUÝN B., ADSERSEN A. and GUDÝKSEN
L., 2006 — Screening of plants used in Danish folk
medicine to treat epilepsy and convulsions. Journal
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MÜLLER A., GANZERA M. and STUPPNER H., 2006
— Analysis of phenolic glycosides and saponins in
Primula elatior and Primula veris (primula root) by
liquid chromatography, evaporative light scattering
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PAULSEN E., CHRÝSTENSEN L.P and ANDERSEN K.E.,
2006 — Miconidin and miconidin methyl ether from
Primula obconica Hance: new allergens in an old
sensitizer. Contact Dermatitis, 55: 203-209.
SUFKA K.J., ROACH J.T., CHAMBLÝSS JR W.G., BROM S.L.,
FELTENSTEÝN M.W., WYANDT C.M. and ZENG L.,
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in the chick social separation-stress procedure. Psy-
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TOKALOV S.V., KÝND B., WOLLENWEBER E. and GUT-
ZEÝT H.E., 2004 — Biological Effects of Epicuticu-
lar Flavonoids from Primula denticulata on Human
Leukemia Cells. J.Agric. Food Chem., 52: 239-245.
TOSUN F., AKYÜZ C., S¸ENER B. and VURAL M., 2005
— The evaluation of plants from Turkey for in vitro
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Received August 6th 2007; accepted February 14th 2008
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Full-text available
  • Dec 2022
In this study, biosynthesis and in vitro phytochemical composition, antibacterial and antioxidant activities of silver nanoparticles were investigated by using aqueous leaf, flower and root extracts of Primula vulgaris (P. vulgaris subsp. vulgaris). The synthesized silver nanoparticles (AgNPs) were confirmed by color conversion and ultraviolet-visible (UV-visible) spectrophotometry. The appearance of a dark brown color and a UV absorption spectrum range at 440 nm confirmed the synthesized silver nanoparticles. The antimicrobial activity of silver nanoparticles synthesized from the leaf of P. vulgaris; S. aureus 25±1, S. epidermidis 20±1, P. aeruginosa 20±1, A. hydrophila 21±1, C. albicans 25±1, C. tropicalis 25±1, C. parapsilosis 22±1 and C. glabrata 20±1 mm zone diameter was determined. The most antimicrobial effect of P. vulgaris leaf aqueous extract; S. aureus 20±1, S. epidermidis 18±1, A.hydrophila 15±1, P. aeruginosa 12±2, C. albicans 18±1, C. glabrata 18±1, C.tropicalis15±2, and C. parapsilosis 15±2 mm zone diameter was revealed. The presence of flavonoids, terpenoids, protein, and carbohydrates was found to be higher in silver nanoparticles synthesized in the flower part of P. vulgaris, according to phytochemical screening. While saponins were detected in P. vulgaris root extracts, tannins and protein were detected in the leaf extract. The flower had the highest total phenolic extract content of the silver nanoparticle (29.08±0 mg GAE/g DW), while the leaf and root had the lowest total phenolic content of 9.06±0.5 and 8.64±3.3 mg GAE/g DW, respectively. The flower had the highest total phenolic extract content of the plant aqueous extracts (25.10±0.2 mg GAE/g DW), while the leaf and root had the lowest (8.28±0.5 and 5.20±0.0 mg GAE/g DW, respectively).The DPPH (1,1-Diphenyl-2-picrylhydrazil) assay was used to assess free radical scavenging activity. The antioxidant activity of AgNPs biosynthesized using P. vulgaris flower extract was 90.6 %, while P. vulgaris flower aqueous extracts were 86.3 %. This can be concluded that silver nanoparticles synthesized using P. vulgaris flower extract are useful in the preparation of pharmacologically useful drugs. Primula vulgaris'ten Sentezlenen Gümüş Nanopartiküllerin Antimikrobiyal ve Antioksidan Potansiyeli ÖZ Bu çalışmada, Primula vulgaris'in (P. vulgaris subsp. vulgaris) sulu yaprak, çiçek ve kök ekstreleri kullanılarak gümüş nanopartiküllerin biyosentezi ve in vitro fitokimyasal bileşimi, antibakteriyel ve antioksidan aktiviteleri araştırılmıştır. Sentezlenen gümüş nanoparçacıklar (AgNP'ler), renk dönüşümü ve ultraviyole-görünür (UV-görünür) spektrofotometrisi ile doğrulandı. Koyu kahverengi rengin görünümü ve 440 nm'de bir UV absorpsiyon spektrum aralığı, sentezlenen gümüş nanoparçacıkları doğruladı. P. vulgaris'in yaprak ekstraktından sentezlenen gümüş nanopartiküldeki en fazla antimikrobiyal etki; S. aureus 25±1, S. epidermidis 20±1, P. aeruginosa 21±1, A. hydrophila 21±1, C. albicans 25±1, C. tropicalis 25±1, C. parapsilosis 22±1 ve C.glabrata 20±1 mm zon çapı ile belirlendi.P. vulgaris yaprak sulu ekstraktın en fazla antimikrobiyal etki; S. aureus 20±1, S. epidermidis 18±1, A.hydrophila 15±1, P. aeruginosa 12±2, C. albicans 18±1, C. glabrata 18±1, C.tropicalis15±2 ve C. parapsilosis 15±2 mm zon çapı ortaya konmuştur. Fitokimyasal taramaya göre P. vulgaris'in çiçek kısmında sentezlenen gümüş nanopartiküllerde flavonoidler, terpenoidler, protein ve karbonhidratların varlığı daha yüksek bulunmuştur. P. vulgaris kök ekstraktlarında saponinler tespit edilirken, yaprak ekstraktında tanen ve protein tespit edildi. P. vulgaris MSU Fen Bil. 1014 yaprak ekstraktından sentezlenen gümüş nanopartiküllerin antimikrobiyal aktivitesinin, P. vulgaris yaprak sulu ekstraktının antimikrobiyal aktivitesinden daha aktif olduğu bulundu. Çiçek, gümüş nanopartikülün en yüksek toplam fenolik ekstrakt içeriğine (29.08±0 mg GAE/g DW) sahipken, yaprak ve kök, sırasıyla 9.06±0.5 ve 8.64±3.3 mg GAE/g DW ile en düşük toplam fenolik içeriğe sahipti. Bitki sulu ekstraktları arasında en yüksek toplam fenolik ekstrakt içeriği çiçekte bulunurken (25.10±0.2 mg GAE/g DW), yaprak ve kök en düşük (sırasıyla 8.28±0.5 ve 5.20±0.0 mg GAE/g DW) bulundu. DPPH (1,1-Difenil-2-pikrilhidrazil) tahlili, serbest radikal süpürme aktivitesini değerlendirmek için kullanıldı. P. vulgaris çiçek özütü kullanılarak biyosentezlenen AgNP'lerin antioksidan aktivitesi %90,6 iken, P.vulgaris çiçek sulu özütleri %86.3'tür. Buradan P. vulgaris çiçek ekstresi kullanılarak sentezlenen gümüş nanopartiküllerin farmakolojik olarak faydalı ilaçların hazırlanmasında faydalı olduğu sonucuna varılabilir.
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... P. vulgaris leaves and root extracts show antibacterial action against E. coli and P. aeruginosa, according to Majid et al. [12]. Saponins and phenolic glycosides are found in the Primula genus [13]. Various species of Primula have been shown to have antioxidant, antibacterial, antimycobacterial, antifungal, cytotoxic, antiviral, antiangiogenic, anti-inflammatory, and antimitotic properties, as well as cytotoxic, antiviral, antiangiogenic, anti-inflammatory, and antimitotic benefits, according to some studies [14][15]. ...
Antimicrobial and Antioxidant Potential of Silver Nanoparticles Synthesized from Primula vulgarisPrimula vulgaris'ten Sentezlenen Gümüş Nanopartiküllerin Antimikrobiyal ve Antioksidan Potansiyeli
Article
  • Sep 2022
In this study, biosynthesis and in vitro phytochemical composition, antibacterial and antioxidant activities of silver nanoparticles were investigated by using aqueous leaf, flower and root extracts of Primula vulgaris (P. vulgaris subsp. vulgaris). The synthesized silver nanoparticles (AgNPs) were confirmed by color conversion and ultraviolet-visible (UV-visible) spectrophotometry. The appearance of a dark brown color and a UV absorption spectrum range at 440 nm confirmed the synthesized silver nanoparticles. The antimicrobial activity of silver nanoparticles synthesized from the leaf of P. vulgaris; S. aureus 25±1, S. epidermidis 20±1, P. aeruginosa 20±1, A. hydrophila 21±1, C. albicans 25±1, C. tropicalis 25±1, C. parapsilosis 22±1 and C. glabrata 20±1 mm zone diameter was determined. The most antimicrobial effect of P. vulgaris leaf aqueous extract; S. aureus 20±1, S. epidermidis 18±1, A.hydrophila 15±1, P. aeruginosa 12±2, C. albicans 18±1, C. glabrata 18±1, C.tropicalis15±2, and C. parapsilosis 15±2 mm zone diameter was revealed. The presence of flavonoids, terpenoids, protein, and carbohydrates was found to be higher in silver nanoparticles synthesized in the flower part of P. vulgaris, according to phytochemical screening. While saponins were detected in P. vulgaris root extracts, tannins and protein were detected in the leaf extract. The flower had the highest total phenolic extract content of the silver nanoparticle (29.08±0 mg GAE/g DW), while the leaf and root had the lowest total phenolic content of 9.06±0.5 and 8.64±3.3 mg GAE/g DW, respectively. The flower had the highest total phenolic extract content of the plant aqueous extracts (25.10±0.2 mg GAE/g DW), while the leaf and root had the lowest (8.28±0.5 and 5.20±0.0 mg GAE/g DW, respectively).The DPPH (1,1-Diphenyl-2-picrylhydrazil) assay was used to assess free radical scavenging activity. The antioxidant activity of AgNPs biosynthesized using P. vulgaris flower extract was 90.6 %, while P. vulgaris flower aqueous extracts were 86.3 %. This can be concluded that silver nanoparticles synthesized using P. vulgaris flower extract are useful in the preparation of pharmacologically useful drugs.
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... The species has been used as a medicinal plant since the Middle Ages, for treatment of gout, headache, and rheumatism [2]. Today, P. veris is widely used for its diverse medicinal effects: secretolytic, expectorant, anti-inflammatory, diuretic, antimicrobial, antifungal, and sedative [3][4][5][6]. Presently, cowslip is less abundant in Europe than in the past, and the sustainable supply of the source material has become more difficult [7,8]. The species is included in the European Red List of Medicinal plants under the category "Least Concern" [9]; however, according to the European Pharmacopoeia, it is still used as a source of Primula roots together with P. elatior [10]. ...
The Establishment of an Ex Situ Collection of Primula veris in Bulgaria
Article
Full-text available
  • Nov 2022
Primula veris is a valuable medicinal plant species with declining populations, protected in Bulgaria by the Biodiversity Act. The present study aimed to increase its extremely low seed germination rate, starting with seeds originating from two Bulgarian populations, and to set up an ex situ field collection. The stimulation effect of three factors was tested in in vivo and in vitro experiments: seeds treated with gibberellic acid (in different concentrations and exposure time), light quality (white, infrared, red, and blue or dark), and cold stratification. The combination of factors resulted in 36 treatment variants in vivo and 8 treatment variants in vitro. No germination was observed in control treatment variants. The highest germinating rate (95%) was noticed in vivo under blue monochromatic light after seed soaking into 0.2% GA3 for 10 h; however, the best results (55% of well-developed seedlings) were observed with a combination of blue light and 0.3% GA3 for 5 h. Seedlings were successfully strengthened in vermiculite in a phytotron, potted in soil and grown in a greenhouse, and then 75 plants were transferred to the field plot, where most of them bloomed at the first vegetation season. These results are intended to serve as a basis for establishing a pilot agriculture of the species.
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The genome sequence of cowslip, Primula veris L.
Article
  • Oct 2024
We present a genome assembly from a specimen of Primula veris (Streptophyta; Magnoliopsida; Ericales; Primulaceae). The genome sequence has a total length of 436.20 megabases. Most of the assembly is scaffolded into 12 chromosomal pseudomolecules. The mitochondrial and plastid genome assemblies have lengths of 326.21 kilobases and 153.48 kilobases, respectively. Gene annotation of this assembly on Ensembl identified 26,584 protein-coding genes.
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The Evaluation of Plants from Turkey for in Vitro. Antimycobacterial Activity
Article
Full-text available
  • Oct 2008
In this research, extracts prepared from plant materials were tested against Mycobacterium tuberculosis. H37Rv (ATCC 27294). Of the 107 plants tested, five exhibited more than 90% inhibition of growth of Mycobacterium tuberculosis., at concentrations lower than 100 µg/ml. Jervin from Veratrum album. L., usnic acid from Usnea barbata. (L.) Mott, and alantolactone from Inula helenium. L. subsp. turcoracemosa. were isolated and their minimum inhibitory concentrations (MICs) were determined as 25, 12.5, and 3.125 µg/ml, respectively.
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Anxiolytic properties of botanical extracts in the chick social separation-stress procedure
Article
Full-text available
  • Feb 2001
The recent growth in sales of natural products labeled as dietary supplements in the United States has renewed scientific interest in the study of the therapeutic effects of multi-component botanical products. This study sought to determine whether botanical extracts derived from the Rutaceae family, Acori graminei, the Magnoliaceae family, Alchemilla vulgaris and Primula veris, which had previously been identified in bioassays as having potential anxiolytic activity, were active in the chick social separation-stress procedure. Eight-day-old chicks received IP injections of test articles 30 min before being tested in the presence of two social companions or in isolation for a 3-min observation period. Dependent measures were: a) latency to adopt a ventral recumbent posture to index sedation, b) number of vocalizations to index separation-distress and c) a composite pain score (comprised of footlift frequency and footlift duration in response to 50 microl of 0.10% formalin injected into the plantar surface of the foot) to index stress-induced analgesia. Proprietal extracts NPS00033 from the Rutaceae plant family and NPS00039 (Relora) from the Magnoliaceae plant family screened positive in this chick model without causing sedation. These results suggest that botanical extracts Relora and NPS00033 may be useful in modulating anxiety states.
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Antimicrobial activity of some endemic plants growing in the Eastern Black Sea Region, Turkey
Article
Full-text available
  • Aug 2006
  • FITOTERAPIA
The Eastern Black Sea Region has an extensive flora because of ample rainfall lasting all year. In this study, antimicrobial effects of 74 crude extracts of 22 endemic plants were investigated. Among the 30 active crude extracts, water-insoluble crude extracts from Betula medwediewii, Heracleum platytaenium, Primula longipes, Anthemis cretica ssp. argaea and Centaurea helenioides were the prominent ones with their MIC values.
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Isolation and characterization of methoxylated flavones in the flowers of Primula veris by liquid chromatography and mass spectrometry
Article
  • Mar 2000
  • J CHROMATOGR A
Characterization of six flavones, which were named substances G1, G2, G3, G4, G5 and G6 according to their RF values in normal-phase thin-layer chromatography, is reported. The pure flavones were purified after maceration with methanol by normal-phase solid-phase extraction, normal-phase medium-pressure liquid chromatography, normal-phase preparative thin-layer chromatography and preparative reversed-phase high-performance liquid chromatography (RP-HPLC). The collected fractions of several isolation steps were analyzed by normal-phase (NP) and RP-HPLC. Detection and identification of the substances G was accomplished by UV detection at 213–216 nm, diode array UV detection, or fluorescence detection (λex=330 nm; λem=440 nm). The molecular mass, the elementary composition, and the structure of the six components was determined by electron-impact high-resolution mass spectrometry (EI-HRMS). Substance G4 was identified as 3′,4′,5′-trimethoxyflavone. The substances G1–G6 were shown to be mono-, di- tri- and pentamethoxyflavones. HPLC–electrospray ionization tandem mass spectrometry (ESI-MS–MS) of the flavones was carried out employing a 150×2 mm I.D. column packed with a 3 μm/100 Å octadecylsilica stationary phase and a mobile phase comprising 1.0% acetic acid in water–acetonitrile (50:50). Comparative RP-HPLC–ESI-MS of the raw methanol extract and the isolated substances G1–G6 proved that the isolated compounds were pure and were not artifacts. Finally, RP-HPLC–ESI-MS–MS was used to identify substances G1–G6 in phytopharmaceutical drugs.
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Chromosome Aberration Assays in Allium. A Report of the U.S. Environmental Protection Agency Gene-Tox Program
Article
  • Dec 1982
  • MUTAT RES-FUND MOL M
The common onion (Allium cepa) is an excellent plant for the assay of chromosome aberrations after chemical treatment. Other species of allium (A. cepa var. proliferum, A. carinatum, A. fistulosum and A. sativum) have also been used but to a much lesser extent. Protocols have been given for using root tips from either bulbs or seeds of Allium cepa to study the cytological end-points, such as chromosome breaks and exchanges, which follow the testing of chemicals in somatic cells. It is considered that both mitotic and meiotic end-points should be used to a greater extent in assaying the cytogenetic effects of a chemical. From a literature survey, 148 chemicals are tabulated that have been assayed in 164 Allium tests for the clastogenic effect. Of the 164 assays which have been carried out, 75 are reported as giving a positive reaction (i.e., causing chromosome aberrations), 49 positive and with a dose response, 1 positive and temperature-related, 9 borderline positive, and 30 negative; 76% of the chemicals gave a definite positive response. It is proposed that the Allium test be included among those tests routinely used for assessing chromosomal damage induced by chemicals.
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Biological Effects of Epicuticular Flavonoids from Primula denticulata on Human Leukemia Cells
Article
  • Feb 2004
The biological effects of epicuticular substances in farinose exudates accumulated on inflorescence shafts and calyces of Primula denticulata on human acute myeloid leukemia cells (HL-60) were analyzed. The crude material possessed little antioxidative capacity but strong cytostatic properties. Some of its known components (5-hydroxyflavone, 2'-hydroxyflavone, 5,2'-dihydroxyflavone, and 5,8-dihydroxyflavone) were further tested to identify the biologically active compounds. The effects of these flavones on cell cycle progression, mitochondrial membrane potential, and reactive oxygen species have been investigated by flow cytometry. The flavonol quercetin was included in the study as reference compound because of its known cytostatic properties and its activity as radical scavenger. Compared to quercetin the flavones induced little apoptosis (up to 40 microM), but despite their low toxicity, the Primula flavonoids possessed strong cytostatic properties even at low concentrations. The cell cycle distribution showed a characteristic time-dependent shift, giving evidence of a generally short-lived effect of the test compounds in the exposed cells. The antioxidative properties quantified according to two different methods correlated with the number of hydroxyl groups. Whereas quercetin strongly affected the mitochondrial membrane potential, none of the Primula flavones showed a comparable effect.
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Lipophilic flavones of Primula veris L. from field cultivation and in vitro cultures
Article
  • Jun 2005
  • PHYTOCHEMISTRY
Ten lipophilic flavones were isolated from the leaves of Primula veris from field cultivation - the newly described 3'-hydroxy-4',5'-dimethoxyflavone and 3'-methoxy-4',5'-methylenedioxyflavone, the previously known from chemical synthesis 3',4'-dimethoxyflavone, 2',5'-dimethoxyflavone, and also flavone, 2'-hydroxyflavone, 2'-methoxyflavone, 3'-methoxyflavone, 3',4',5'-trimethoxyflavone and 5,6,2',6'-tetramethoxyflavone (zapotin) which were previously known from plants. The same flavones were found in the leaves of P. veris obtained by in vitro propagation. The structural assignments were derived from (1)H NMR, (13)C NMR, EIMS and UV spectral data and the influence of B-ring oxygen substituents on the C-2, C-3 and H-3 NMR resonances in flavones unsubstituted in the A ring is taken into consideration.
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Screening of plants used in Danish folk medicine to treat epilepsy and convulsions
Article
  • May 2006
  • J ETHNOPHARMACOL
Aqueous and ethanolic extracts of 42 plants used in Danish folk medicine for the treatment of epilepsy and convulsions, or for inducing sedation, were tested for affinity to the GABA(A)-benzodiazepine receptor in the flumazenil-binding assay. Ethanolic extracts of leaves of Primula elatior and Primula veris and aerial parts of Tanacetum parthenium exhibited good, dose-dependent affinity.
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Analysis of phenolic glycosides and saponins in Primula elatior and Primula veris (primula root) by liquid chromatography, evaporative light scattering detection and mass spectrometry
Article
  • Apr 2006
  • J CHROMATOGR A
This paper describes the first liquid chromatographic method suitable for the simultaneous determination of bioactive compounds, saponins and phenolic glycosides, present in Primula elatior and Primula veris, including the NMR data of primulaverin and primeverin. Optimum separations were obtained with a Synergi 4mum Fusion RP 80A column, using 0.025% TFA in water and 5% acetonitrile in methanol as mobile phase. Saponins were detected by evaporative light scattering detection (ELSD), whereas the phenolic glycosides were monitored by UV at 210 nm. The method was validated for repeatability (sigma(rel) <or=4.5%), precision (intra- and inter-day variation <or=5.0%), accuracy (recovery >or=97.1%) and sensitivity (LOD <or=22 ng (UV) and <or=38 ng (ELSD) on-column, respectively). LC-MS experiments in negative APCI mode allowed a final peak assignment. Both Primula species could easily be differentiated by their saponin pattern. The total saponin content was highest in P. veris roots (max. 14.9%), the aerial parts or P. elatior contained significantly less amounts; primeverin (0.64-1.42%) showed to be the most dominant phenolic glycoside.
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