Primula Denticulata Research Articles

Development of genome-wide microsatellites from Primula denticulata and their utility in different species of the genus

The genus Primula (Primulaceae) harbours habitat-sensitive species. It is a valuable tool for monitoring and assessing the overall state of the ecosystem as it can serve as a reliable indicator of ecological health. Primula denticulata Sm. is one of the most prevalent Primrose species found in the Indian Himalayan region with a wide extent of distribution from 1500 m to 3500 m altitude. Considering the ecological importance of Primulas, it is imperative to develop microsatellite markers to study genetic variability which can offer valuable insights into the adaptive strategies of the plants in response to changing environmental conditions. In this study, we sequenced the P. denticulata genome by utilizing Illumina paired-end sequencing technology at ∼15X coverage. 27,196 sequences harbouring 31,584 microsatellite loci were identified. 100 primer pairs were synthesized and tested for amplification. Impressively, 84 of these primers exhibited robust amplification, and 64 of these displayed polymorphism. Primer pairs from trinucleotide motifs were identified as more polymorphic. To assess the broader applicability of these primers, we also tested cross-species transferability on 14 different Primula species representing different sections of the genus that were collected from different parts of Himalaya. Notably, one SSR locus, Next Generation Sequencing Primula_5 (NGSPr_5) was able to produce bands in all the species showing 100 % transferability. Primula atrodentata showed a maximum (88.23 %) while P. glabra exhibited minimum (19.60 %) cross-transferability. The UPGMA dendrogram clustered different sections of sub-genus Aleuritia, and species from subgenera Sphondylia and Auganthus separately into two major groups. The groupings are in accordance with the known relationship between the different species from the genus.

In- Vitro antioxidant activity of Primula denticulata aerial parts extracts

Abstract- In this study, we identify the anti-oxidant impact of ethanolic and aqueous extract of aerial parts of Primula denticulata and to evaluate the anti-oxidant effect of ethanolic and aqueous extract and compare to it with ascorbic acid and BHT by DPPH Radical Scavenging Assay methods because there is no scientific data available on the anti-oxidant activity of plant. The current investigation also showed that Primula denticulata aerial portions contain a variety of secondary metabolites. These phytochemicals may be a significant source of pharmacological compounds, meaning that the plant species may have enormous potential for use as a treatment for a range of chronic illnesses. The species' crude extract exhibits encouraging antioxidant potential as well, supporting the traditional use of this plant with scientific evidence.

Flower phenological events and duration pattern is influenced by temperature and elevation in Dhauladhar mountain range of Lesser Himalaya

Studying phenology is undeniably one of the most effective ways to monitor and perceive how a particular plant species interact and respond to varying environmental conditions. In this study, the shifts in flowering phenological events of 24 herbaceous species with the changing elevation and temperature were observed in Dhauladhar Mountain range of Lesser Himalaya. Five permanent plots (20 × 20 m2) were established from 2,000 m to 4,000 m elevation at each rising 500 m distance for the documentation of herbaceous flora. Abundance-based documentation of flowering phenological events (onset of flowering duration [OFD] ≥ 10% to 30%, the peak of flowering duration [PFD] > 30% to 80%, the end of flowering duration [EFD] > 80% and total flowering duration [FD] as the difference of end and onset of flowering DOY) was done in each plot. The onset, the peak, and the end of the flowering day of the year (DOY) were recorded at 20%, 55%, and 90% floral abundance, respectively, from 1st of January. Upon analyses, the Linear-mixed effects model depicted significant differences in the flowering phenological events with elevation and soil temperature. The onset of flowering DOY and peak of the flowering DOY varied significantly with soil temperature gradient, while the end of flowering DOY showed non-significant interaction. The linear model of ANOVA demonstrated that OFD, PFD, EFD and FD varied significantly with elevation at p < 0.001. FD increased with elevation for most of the species, whereas two species, Aquilegia pubiflora and Primula denticulata, showed early incidences of flowering compared to other species. Temperature played an imperative role in influencing the day of initiation and duration of these phenological events. High plasticity of flowering phenological events is advantageous in high elevation regions where pollinators are scarcely present. Therefore, documenting shifting incidences of flowering phenological events would help the researcher to effectively predict climate change effects on alpine communities in near future.

Habitat selection of Himalayan Musk Deer Moschus leucogaster (Mammalia: Artiodactyla: Moschidae) with respect to biophysical attributes in Annapurna Conservation Area of Nepal

Himalayan or White-bellied Musk Deer Moschus leucogaster, an IUCN indexed endangered species, is distributed in isolated pockets in the Himalaya. The deer population is decreasing owing to several pressures that include habitat loss and fragmentation, and poaching. It is essential to identify preferred habitat characteristics to support appropriate management strategies for conserving this endangered species. This study was carried out in the Nysheang basin of Annapurna Conservation Area of Nepal to identify habitats preferred by the musk deer. Habitat field parameters were collected using transect surveys. To analyze vegetation use and availability, nested quadrate plots size 20 m2 were established. Ivlev’s electivity index (IV) (-1 to +1) was employed to determine habitat preference, and one-way ANOVA (F) and chi-square tests (χ2) were used to examine different habitat parameters. Similarly, the importance value index (IVI) of the vegetation was calculated. Our results showed that the Himalayan Musk Deer strongly preferred habitats at 3601–3800 m altitude (IV= 0.3, F= 4.58, P &lt;0.05), with 21–30º slope (IV= 0.2, F= 4.14, P &lt;0.05), 26–50 % crown cover (IV= 0.25, F= 4.45, P &lt;0.05), 26–50 % ground cover (IV= 0.15, F= 4.13, P &lt;0.05), and mixed forest (IV= 0.29, χ2= 28.82, df= 3, p &lt;0.001). Among the trees, Abies spectabilis (IVI= 74.87, IV= 0.035) and Rhododendron arboretum (IVI= 55.41, IV= 0.02) were the most preferred, while Rhododendron lepidotum, Cassiope fastigiata (IV= 0.35) and Berberis aristata (IV= 0.25) were the most preferred shrubs, and Primula denticulata (IV= 0.87) and Primula rotundifolia (IV= 0.31) were the most preferred herbs. These preferred habitat conditions should be maintained and conserved to sustain a viable population of deer in the study area. Further studies will be required to assess the effects of climate change on habitat suitability.

Determination of the optimum extraction regime of reducing compounds and flavonoids of Primula denticulata Smith leaves by a dispersion analysis

Herbal medicines are widely used in the complex treatment of various diseases. Therefore, theoretical and practical interest is the in-depth study of drumstick primrose (Primula denticulata Smith). The study aimed to determine the optimal extraction mode of flavonoids and reducing compounds of drumstick primrose leaves. The concentration of ethanol, the ratio of raw materials and extractant, and extraction method were studied by dispersion analysis. This allowed reducing the number of experiments from 64 to 16. To obtain the alcohol extract of drumstick primrose leaves with the highest content of reducing compounds and flavonoids, found that maceration is the optimal method of extraction, the ratio of raw materials to extractant should be 1 to 5 and 40% ethanol is the most appropriate extractant.

Traditional practices of the ethnoveterinary plants in the Kaghan Valley, Western Himalayas-Pakistan

Introduction: Indigenous people in a far-flung mountainous area without basic facilities, mainly rely on medicinal plants to cope with various veterinary health problems. Objective: The present study was carried out to explore the traditional knowledge of ethnoveterinary practices in Kaghan Valley, district Mansehra, Western Himalayas-Pakistan. Method: Ethnoveterinary data were collected between February to October 2014 from nine villages of the Kaghan Valley by involving 80 local people include traditional healers using a semi-structured interview. Results: A sum of 41 plant taxa of ethnoveterinary medicinal plants was documented for treating livestock ailments. Out of which, herbaceous plants were recorded with high percentage (27 species, 65.8 %). Most of the species were used to treat gastrointestinal diseases (12 taxa), followed by health improvement (7 taxa). The widely used part of plants for livestock aliments was the whole plant (9 species) followed by leaves (7 species), and preparation were paste (18 species) followed by powder with 10 species. The highest used values were recorded for Arisaema costatum (0.82), Primula denticulata (0.76), and high relative frequency citations for Berberis lyceum and Dryopteris ramosa with 0.37 each. Among the plant species Skimmia laureola, Thymus linearis and Phytolacca latbenia were among the taxa with cent fidelity level. Conclusion: The flora used in traditional remedies of the valley was found mostly endemic due to excessive utilization. Thus, further chemical investigation, better utilization and conservation of indigenous use of the reported species should be considered for future work.

Characterization of 30 microsatellite markers for distylous Primula denticulata (Primulaceae) using HiSeq sequencing.

Primula denticulata exhibits considerable variation in floral morphology and flowering phenology along elevational gradients in SW China. We isolated 30 microsatellite markers from P. denticulata to facilitate further investigation of population genetics and floral evolution in this species. We used the HiSeq X-Ten sequencing system to develop a set of markers, and measured polymorphism and genetic diversity in a sample of 72 individuals from three natural populations of P. denticulata subsp. denticulata. The markers displayed relatively high polymorphism, with the number of alleles ranging from two to seven (mean = 3.567). The observed and expected heterozygosity ranged from 0 to 1.000 and 0.041 to 0.702, respectively. Twenty-eight of the loci were also successfully amplified in P. denticulata subsp. sinodenticulata. The microsatellite markers we have identified will provide valuable tools for investigations of the population genetic structure, mating systems and phylogeography of the P. denticulata complex, and will help to address questions concerning the ecological and genetic mechanisms responsible for the evolution of reproductive traits in the species.

ВМІСТ САПОНІНІВ У ЛИСТКАХ І КОРЕНЕВИЩАХ З КОРЕНЯМИ КУЛЬТИВОВАНИХ ВИДІВ РОДУ PRIMULA L.

Вступ. У світoвій флoрі рід Primula L. родини Primulaceae є одним із найчисленіших та oб’єднує понад 600 видів одно- і багаторічних рослин. На території України зростає 9 видів роду Примула. У джерелах літератури є інформація про цілющі властивості примули скельної (Primula saxatilis Коm.) та примули дрібнозубчастої (Primula denticulatа Smith.), які використовують тільки в народній медицині. Водночас у наукових джерелах літератури недостатньо відомостей про їх хімічний склад.&#x0D; Мета дослідження – визначити вміст сапонінів у листках і кореневищах з коренями декоративних видів роду Примула – примули скельної та примули дрібнозубчастої.&#x0D; Метoди дoслідження. З метoю виявлення сапонінів у дoсліджуваній сирoвині прoвoдили загальнoприйняті якісні реакції, для чoгo пoпередньo гoтували витяжки. Водні витяжки використовували для проведення проби на піноутворення та осадових реакцій. Визначали також хімічну природу сапонінів у досліджуваних об’єктах. Кількісний вміст сапонінів у перерахунку на есцин визначали спектрофотометричним методом на спектрoфoтoметрі Lambda 25 Perkin Elmer (США) при довжині хвилі 381 нм.&#x0D; Результати й обговорення. У результаті прoведених досліджень встановлено, що вміст сапонінів у примули дрібнозубчастої листках в 1,4 раза більший, ніж у примули скельної листках, і становив (0,61±0,03) %, вміст сапонінів у примули дрібнозубчастої кореневищах з коренями в 1,2 раза вищий, ніж у примули скельної кореневищах з коренями, і складав (1,79±0,06) %.&#x0D; Висновки. Проведено фітохімічний аналіз листків і кореневищ з коренями примули скельної та примули дрібнозубчастої, експериментально доведено наявність сапонінів тритерпенового ряду. Вміст сапонінів у примули скельної листках і кореневищах з коренями становив (0,45±0,02) та (1,54±0,07) %, у примули дрібнозубчастої − (0,61±0,03) і (1,79±0,06) % відповідно.

Investigation of morphological and anatomical underground parts structure of genus Primula L. Cultivated species

Investigation of morphological and anatomical underground parts structure of genus Primula L. Cultivated species

Effects of cold-storage duration on growth and floral development of dormant plants ofPrimula denticulatasubsp.sinodenticulatain water culture

Effects of cold-storage duration on growth and floral development of dormant plants of<i>Primula denticulata</i>subsp.<i>sinodenticulata</i>in water culture

ANTI-OXIDANT AND ANTI-DIABETIC ACTIVITIES OF ETHANOLIC EXTRACT OF Primula denticulata FLOWERS

The present investigation was carried out to focus on the anti-oxidant and anti-diabetic effect of ethanolic extract of flowers of Primula denticulata by DPPH, ferrous chelating, reducing power assay and streptozotocin induced diabetes respectively. Dose selection was made on the basis of acute oral toxicity study (100mg/kg, 200mg/kg, 400mg/kg bodyweight) as per OECD guidelines 423. The blood glucose levels were measured by using blood glucose measuring strips based on glucose-oxidase method. The extract showed significant anti-oxidant and anti-diabetic activity when compared with standard drug. Keywords: Primula denticulata, anti-oxidant and anti-diabetic activity.

In Vitro Antioxidant, Antibacterial Activity and Phytochemical Studies of Primula Denticulata An Important Medicinal Plant of Kashmir Himalaya

The present study was designed to evaluate the phytochemical composition, antimicrobial and radical-scavenging activities of ethanolic extract of Primula denticulata- an important medicinal herb of Kashmir Himalaya. Phytochemical study was performed by using various standard phytochemical methods. Free radical scavenging activities of the extract was assessed by employing different in vitro assays such as DPPH free radical scavenging assay, hydrogen peroxide scavenging activity and lipid per oxidation assay. Calf thymus DNA was also monitored by TBARS formation. The results were compared with standard antioxidant (?-tocopherol). Antibacterial activity of the extract was determined by agar well diffusion method. DPPH free radical scavenging assay and hydrogen peroxide scavenging activity revealed plant extract to be an active radical scavenger. P.denticulata extract also dose dependently inhibited the MDA formation or lipid per oxidation and as such might intercept the free radical chain of oxidation. The leaf extract also prevents calf thymus DNA from oxidative damage induced by hydroxyl radical generated by FeSO4 and H2O2 in Fenton reaction. The hydroxyl radical quenching ability of polyphenolic compounds of Primula denticulata could be responsible for the protection against oxidative damage to DNA. The ethanolic extract showed anti-microbial activity which was visible as the zones of inhibition formed in the different cultures of Gram positive and Gram negative bacteria as well as in case of fungal cultures. Among all the maximum activity was seen for Escherichia coli with inhibition zone diameter of 23.53 3.71mm followed by Klebsiella pneumoniae with inhibition zone diameter of 20.74 3.33mm. These findings provide evidence that the considered plant possesses antimicrobial and antioxidant properties and may act as possible antioxidant for biological systems susceptible to free radical-mediated reactions.

Altitudinal Variation In Some Phytochemical Constituents And Stomatal Traits Of Primula Denticulata

Due to extensive medicinal importance and wide distributional range of Primula denticulata Sm., locally known as Doker-neej (Kashmiri) of family Primulaceae, in Kashmir valley, the present investigation was carried out to study the variations in phenology, stomatal traits and content of phytochemicals of the species in relation to altitude. During the present investigation, trends in the stomatal traits which include: stomatal density, length and breadth of guard cells and potential conductance index (PCI) along an altitudinal gradient were analyzed by using standard methods. Samples from different populations with varying altitudes were also subjected to qualitative and quantitative estimation for the presence of various phytochemicals by using various spectrophotometric techniques.It was observed that mean values of the analyzed stomatal traits in the four populations varied significantly (p?0.05) across the populations. Phytochemicals such as phenols, tannins, alkaloids and saponins were present in all the sampled populations but their quantity varied significantly across the populations. This assessing of phytochemical content of plants at varying altitudes can help to select elite genotype and reflect the best suited altitude for commercial cultivation of the species as these phytochemicals are considered as the basis for their medicinal activity.

A study of medicinal plants used as ethnoveterinary: Harnessing potential phytotherapy in Bheri, District Muzaffarabad (Pakistan)

Ethnopharmacological relevanceMedicinal plants are utilized for handling health care system and in preventing a variety of diseases. A survey was conducted to document the rapidly disappearing traditional knowledge of medicinal plants in union council Bheri, District Muzaffarabad, Azad Kashmir, Pakistan. Materials and methodsQuestionnaire format was used to collect the medicinal uses of plants. The 180 informants were interviewed from six villages in total, 30 from each village (20 male and 10 female) regarding the ethnoveterinary uses of plants in several ailments. For the reliability of ethnoveterinary knowledge, the informant consensus factor (FIC), and fidelity level (FL) were calculated and the literature cited was surveyed. The medicinal information was gathered from local inhabitants, healers, shepherds and old men and women of different age groups. ResultsA total of 24 medicinal plant species used as ethnoveterinary were found belonging to 22 genera and 19 families. The most dominant family was Polygonaceae (3 species) followed by Araceae, Asteraceae, Lamiaceae each with 2 species and remaining families having one species. The important medicinal plant species showed the highest fidelity level (FL) such as: Rumex nepalensis, Primula denticulata, (100%) used for dysuria, red urination, Skimmia laureola (100%), Swertia paniculata (99%), and Angelica glauca (97%), used for ague, cold, shivering, gastric ailments, Melia azedarach (100%), used to reduce intestinal worm load in cattle showing the conformity of knowledge on these species. Highest FIC was recorded for foot and mouth diseases and ectoparasite (1) followed by ague (0.98) and dysuria (0.99) depicting that a few species were used to cure various animals׳ ailments. ConclusionsThe findings of the research revealed that merely a few species are used as ethnoveterinary medicine supported by pharmacology study. Due to anthropogenic pressure the extinction of each species from the areas could result in disappearing knowledge regarding century׳s old traditional methods of curing diseases from these plant species.

Unreduced 3x gamete formation of allotriploid hybrid derived from the cross of Primula denticulata (4x) × P. rosea (2x) as a causal factor for producing pentaploid hybrids in the backcross with pollen of tetraploid P. denticulata

A triploid hybrid, which was obtained from interspecific crosses between tetraploid Primula denticulata (2n = 4x = 44) and P. rosea (2n = 2x = 22), successfully produced 11 plants by backcrossing with pollen of tetraploid P. denticulata. Analysis of ploidy level using flow cytometry and chromosome counting in the 11 BC1 plants revealed that all progeny had much larger DNA contents and chromosome number than both parents. In this triploid-tetraploid (3x–4x) crossing, progeny was predominantly true or near pentaploid presumably produced by the fertilization between true or near triploid female gamete produced from triploid hybrid and diploid pollen of tetraploid P. denticulata. These results suggest that unreduced (3x) or near triploid female gametes were partially produced by single step meiosis, either first-division restitution or second-division restitution process. The zygotes formed by the fertilization between true or near triploid egg produced by single step meiosis in triploid hybrid and diploid pollen produced by normal meiosis of tetraploid P. denticulata might be the only survivors in embryogenesis.

Plant diversity in alpine pasture of Talra wildlife sanctuary of district Shimla, Himachal Pradesh

Studies to evaluate the plant diversity and growth forms in alpine pasture of Talra Wildlife Sanctuary of District Shimla, Himachal Pradesh were carried out during August, 2003. Total 75 species of plant, comprising of 7 grasses, 4 sedges, 4 leguminous forbs and 60 non-leguminous forbs were recorded from the area. On the basis of Importance Value Index (IVI). Sibbaldia cuneata O. Ktze., was found to be the dominant species followed by Geum elatum Wall ex D. Don., Ligularia amplexicaulis DC., Primula denticulata Smith and Saxifraga parnassifolia D. Don. The distribution of all the plant species was contagious. Index of dominance was 0.036 and index of diversity 5.386. The contribution of tall forbs, short forbs and cushioned and spreading forbs in the alpine pasture was 29.33%, 54.66% and 16.0% respectively.

Inter-sectional Hybrids with Various Ploidy Levels between Primula denticulata and Three Varieties of P. modesta

Reciprocal crossing was carried out between tetraploid Primula denticulata, section Denticulata and three taxa of P. modesta, diploid P. modesta var. fauriae, diploid P. modesta var. samanimontana and tetraploid P. modesta var. matsumurae, section Aleuritia. Inter-sectional hybrids were successfully obtained in all the reciprocal crosses except when P. modesta var. samanimontana was crossed with the pollen of P. denticulata. The number of hybrids obtained by using P. modesta var. fauriae as maternal parent was extremely low, whereas a large number of hybrids was obtained in the crosses when tetraploid species, P. denticulata and P. modesta var. matsumurae, were used as maternal parent. In all the cross combinations using P. denticulata as maternal parent, both legitimate and illegitimate crosses yielded germinable hybrid seeds. This suggested that in tetraploid P. denticulata heterostylous incompatibility might have decreased or had been lost at least on the female side. In most crosses, the DNA content of the hybrid was intermediate between that of the parents, except for the cross of P. modesta var. fauriae × P. denticulata, where the tetraploid hybrids were mainly produced due to the fertilization of the unreduced female gametes of P. modesta var. fauriae. These results suggest that in most of the zygotes with a lower ratio of female genome than male, development was inhibited in these inter-specific hybridizations.

Variation of ploidy level in inter-section hybrids obtained by reciprocal crosses between tetraploid Primula denticulata (2n = 4x = 44) and diploid P. rosea (2n = 2x = 22)

SummaryInter-section hybrids were successfully obtained from reciprocal crosses between tetraploid Primula denticulata (section Denticulata) and diploid P. rosea (section Oreophlomis) by applying ovary culture 4 weeks after pollination. Differences in ploidy levels were found in these hybrids by flow cytometric analysis and by counting chromosome numbers. When P. denticulata was used as maternal parent, all the hybrids were triploid, with two genomes of P. denticulata and one genome of P. rosea. In contrast, when P. rosea was used as maternal parent, two genome-type hybrids were obtained: one was a triploid hybrid, with the same genome combinations as those obtained when P. denticulata was used as maternal parent, and the other type was a tetraploid hybrid with two genomes of each species. These results suggest that unreduced (2x) gametes were formed only in the female side of P. rosea.

Triterpenoid Saponins fromPrimula denticulata

Two triterpene glycosides denticin ( 1) and denticulatin ( 2) have been isolated from the whole plant of PRIMULA DENTICULATA Sm. and their structures were established by spectral and chemical studies as 3beta-{ O-beta- D-glucopyranosyl-(1-->4)-[ O-beta- D-xylopyranosyl-(1-->2)]- O-(beta- D-glucopyranosyl-(1-->4)-[ O-beta- D-glucopyranosyl-(1-->2)]-alpha- L-arabinopyranosyloxy}-16alpha-hydroxy-13beta, 28-epoxyolean-30-al and 3beta-{ O-beta- D-glucopyranosyl-(1-->2)- O-beta- D-glucopyranosyl-(1-->4)-[ O-beta- D-xylopyranosyl-(1-->2)]- O-beta- D-glucopyranosyl-(1-->4)-[ O-beta- D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyloxy}-16alpha-hydroxy-13beta,28-epoxyolean-30-al, respectively.

Karyomorphological studies in some Primula species of Nepal Himalaya.

Karyotype analysis were made using root tip, leaf tip, petal tip as well as pollen mitosis in 13 species of Nepal himalayas. The chromosome number of Primula atrodentata W. W. Sm. (2n=22); Primula deuteronana Craib (2n=22); Primula edgeworthii Pax (2n=22); Primula erosa Wall. (2n=22, 24); Primula irregularis Craib (2n=22); Primula rotundifolia Wall. (2n=22) are reported here for the first time. Genetical diversity has been recorded in Primula denticulata Sm. with having variable chromosome numbers of 32, 34, and 44. The chromosome number of Primula elliptica Royle is observed as 2n=24.Data on the chromosome numbers of Primula gracilipes Craib (2n=22); Primula malacoides Franch. (2n=18); Primula petiolaris Wall. (2n=22); Primula reticulata Wall. (2n=22); Primula sikkimensis Hook. f. (2n=22), reported in previous references have been confirmed.The basic number 11 is predominating in this genus studied. Critical notes are made in the karyotype characteristics and its interrelationship among species.