Longevity and germination of Juniperus communis L. pollen after storage.

Pollen storage belongs among the most important activities associated with pollen handling. It overcomes the differences in pollen shedding and ovule receptivity during controlled pollination experiments. It is especially important for species like common juniper (Juniperus communis L.) with an extremely low quality of seeds due to pollination failure. Additionally, it is a substantial part of germplasm preservation programmes in pollen banks. In the present paper, the effect of short-term storage of pollen was studied using pollen samples from five shrubs in an in vitro germination test. Two temperature regimes were tested. The pollen viability of freshly collected pollen varied considerably between individual shrubs, exhibiting 67.3-88.6% germination rate and 248.0-367.3 µm of pollen tubes. Storage at + 4 °C for four months was accompanied by a profound decline in pollen viability. The germination percentage was reduced to 49.2-75.2% and the pollen tube length to 32.5-69.0%, depending on individual shrubs. The corresponding decline in pollen viability characteristics during storage at - 20 °C was only negligible in two of the tested shrubs. In the remaining three shrub samples, an increase in germination percentage was observed. Pollen tube growth responded more sensitively to freezing, but, on average, the decrease in length was lower than that at + 4 °C. The rate of reduction in pollen tube length varied between 11.5 and 45.4%. Cytological events accompanying in vitro germination of freezer-stored pollen exhibited some delay in releasing the exine from pollen grains during the early stages of germination as compared with freshly collected pollen. In conclusion, short-term storage of the common juniper pollen in a freezer is better for the preservation of its viability than storage at + 4 °C.

Mass propagation of Juniperus procera Hoechst. Ex Endl. From seedling and screening of bioactive compounds in shoot and callus extract.

BACKGROUND: Juniperus procera Hoechst. ex Endl. is a medicinal tree in Saudi Arabia, primarily in the Enemas region, but it is locally threatened due to die-back disease and difficulties regarding seed reproduction (seed dormancy and underdeveloped embryonic anatomy, and germination rate < 40%). Hence, the alternative methods for reproduction of Juniperus procera are really needed for conservation and getting mass propagation for pharmaceutical uses. RESULTS: In this manuscript, we articulated the successful in vitro shoot multiplication and callus induction of J. procera by using young seedling as explants and detected an important antibacterial and antitumor product. Explants were grown on different types of media with the supplement of different combinations of Plant Growth Regulators (PGRs) at different concentrations. The best media for shoot multiplication was Woody Plant Media (WPM) supplemented with PGRs (0.5 µM of IAA and 0.5 µM BAP or 0.5 µM IBA and 0.5 µM BAP). Whereas for callus induction and formation Woody Plant Media (WPM) with the addition of PGRs (0.5 µM 2,4-D and 0.5 µM BAP) was better than the Chu Basal Salt Mixture (N6), Gamborg's B-5 Basal Medium (B5), and Murashige and Skoog media. The possibility of multiplication of J. procera in vitro creates significant advantages to overcome the difficulties of seeds dormancy for the reproduction of plants, conservation of trees, and getting mass propagation material for pharmaceutical studies. The shoot and callus extract of J. procera was detected using gas chromatography-mass spectrometry analysis and revealed more than 20 compounds related to secondary metabolites, which contained antibacterial and antitumor agents, such as ferruginol, Retinol, and Quinolone as well as confirmed by Direct Analysis in Real Time, Time of Flight Mass Spectrometry (DART-ToF-MS). Podophyllotoxin (PTOX) was detected in callus material by HPLC with sigma standard and confirmed by DART-ToF-MS and UV spectra. CONCLUSION: We successfully conducted in vitro shoot multiplication and callus induction from J. procera seedlings using WPM and a different combination of PGRs and, detected an important antibacterial and antitumor product such as ferruginol and podophyllotoxin. According to our findings, J. procera has become a new natural source of novel bioactive compounds.

Variations in leaf traits of Juniperus przewalskii from an extremely arid and cold environment.

How leaf traits vary with environmental and climatic variables in cold and arid environments is an essential issue in environmental ecology. Here, we analyzed the variations in leaf nitrogen (N) and phosphorus (P) stoichiometry and leaf dry matter content (LDMC) in Qilian juniper (Juniperus przewalskii Kom.) growing in 14 environmentally different plots on the northeastern Tibetan Plateau. The results showed that the N and P concentrations, N:P ratio and LDMC of Qilian juniper were 10.89 mg.g-1, 1.04 mg.g-1, 10.80 and 483.06 mg.g-1, respectively. The spatial coefficients of the variations in leaf N and P stoichiometry were significantly higher than the seasonal ones, and the correlations of leaf N and P concentrations with spatial variables were stronger than their correlations with the season. During the growing season, only the leaf N concentration and N:P ratio significantly increased. Soil nutrients were highly positively significantly correlated with leaf P concentrations but negatively correlated with the N:P ratio and LDMC. However, leaf N concentrations showed no significant correlations with soil nutrients. We suggest that the effects of temperature on the N concentration and LDMC were stronger than the effects of drought, while those on the P concentration and N:P ratio were weaker. Drought reduced leaf N and P concentrations and increased the N:P ratio and LDMC. In the arid region, with an increasing mean annual temperature (MAT), leaf N concentration significantly decreased, and LDMC significantly increased. In the semi-arid region, as MAT increased, leaf N and P concentrations significantly increased and LDMC and the N:P ratio significantly decreased. These opposite results supported the growth rate hypothesis that plant N and P concentrations increase while the N:P ratio and LDMC decrease as the growth rate increases.

Phenolic profile and in vitro bioactive potential of Saharan Juniperus phoenicea L. and Cotula cinerea (Del) growing in Algeria.

The aim of this study was to characterize the individual phenolic profile and antioxidant, anti-inflammatory, cytotoxic, and antimicrobial activities of hydroethanolic and infusion extracts prepared from Algerian Saharan Juniperus phoenicea L. and Cotula cinerea (Del). The phenolic profile was determined using a liquid chromatograph coupled to a diode array detector and an electrospray ionization mass spectrometer (LC-DAD-ESI/MS). A total of thirteen and nine individual phenolic compounds were identified in J. phoenicea and C. cinerea, respectively. 3-p-Coumaroylquinic acid, quercetin- and myricetin-O-pentoside were the major compounds present in J. phoenicea; on the other hand, C. cinerea presented luteolin-7-O-glucoside, luteolin-O-malonylhexoside, and 5-O-caffeoylquinic acid as the main molecules. In general, all samples exhibited interesting antioxidant activity when compared to the standard Trolox, but J. phoenicea extracts presented the highest bioactivity. Likewise, all the samples exhibited anti-inflammatory activity; thus J. phoenicea hydroethanolic extracts showed the highest potential (88 ± 8 µg mL-1). In addition, their cytotoxicity was evaluated towards a panel of four selected cell lines (HeLa, NCI-H460, MCF-7 and HepG2), and all the extracts showed cytotoxic effects, with J. phoenicea extracts being the most effective. The in vitro antimicrobial activity of the plant extracts was moderate, Gram-positive bacteria thus being more sensitive than the Gram-negative strains (MIC values between 5 and 20 mg mL-1). The present work suggests that J. phoenicea and C. cinerea are sources of bioactive ingredients with potential use in the food and pharmaceutical industries.

Juniperus chinensis Fruits Attenuate Oxazolone- and 2,4-Dinitrochlorobenzene-Induced Atopic Dermatitis Symptoms in Mice.

Juniperus chinensis, commonly Chinese juniper, has been used for treating inflammatory diseases. This study aimed to investigate anti-atopic dermatitis (AD) effects of standardized J. chinensis fruits extract on murine oxazolone- and 2,4-dinitrochlorobenzene (DNCB)-induced models of AD. Ear swelling, epidermis thickening, and eosinophils infiltration in the oxazolone-mediated dermatitis of BALB/c mice were significantly reduced upon topical application of J. chinensis fruits 95% EtOH extract (JCE). Besides, transdermal administration of JCE to SKH-1 hairless mice inhibited the development of DNCB-induced AD-like skin lesions by suppressing transepidermal water loss and improving skin hydration. Decreased total serum immunoglobulin E (IgE) and interleukin (IL)-4 levels could be observed in atopic dorsal skin samples of JCE-treated group. According to the phytochemical analysis, JCE was found to contain isoscutellarein-7-O-ß-D-xyloside, cupressuflavone, and amentoflavone as main compounds. Therapeutic attempts with the J. chinensis fruits might be useful in the treatment of AD and related skin inflammatory diseases.

Production of Podophyllotoxin by Plant Tissue Cultures of Juniperus virginiana.

Plant tissue cultures are a potential source of secondary metabolites. However, their production, when compared with intact plants, is usually lower. Phenylalanine, a biogenetic precursor of podophyllotoxin, was used to stimulate podophyllotoxin production in callus and suspension cultures of Juniperus virginiana L. The best phenylalanine effect on podophyllotoxin production was manifested in three-years-old callus cultures after a 21-days application of a 10 mmol/L concentration. A podophyllotoxin content of 0.15 mg/g DW was determined, which was about 400% higher in comparison with the control. The maximum content (0.48 mg/g DW) in newly derived suspension cultures (the 4' passage) was induced by 14-days application of a I mmol/L concentration; this was about 243% higher than the control. In one-year-old suspension cultures the highest podophyllotoxin content (0.56 mg/g DW) was recorded also after 14-days application of a I mmol/L concentration; this was about 211% higher than in the control cultures.

Western Juniper Management: Assessing Strategies for Improving Greater Sage-grouse Habitat and Rangeland Productivity.

Western juniper (Juniperus occidentalis subsp. occidentalis) range expansion into sagebrush steppe ecosystems has affected both native wildlife and economic livelihoods across western North America. The potential listing of the greater sage-grouse (Centrocercus urophasianus) under the U.S. Endangered Species Act has spurred a decade of juniper removal efforts, yet limited research has evaluated program effectiveness. We used a multi-objective spatially explicit model to identify optimal juniper removal sites in Northeastern California across weighted goals for ecological (sage-grouse habitat) and economic (cattle forage production) benefits. We also extended the analysis through alternative case scenarios that tested the effects of coordination among federal agencies, budgetary constraints, and the use of fire as a juniper treatment method. We found that sage-grouse conservation and forage production goals are somewhat complementary, but the extent of complementary benefits strongly depends on spatial factors and management approaches. Certain management actions substantially increase achievable benefits, including agency coordination and the use of prescribed burns to remove juniper. Critically, our results indicate that juniper management strategies designed to increase cattle forage do not necessarily achieve measurable sage-grouse benefits, underscoring the need for program evaluation and monitoring.

Climate change and land management in the rangelands of central Oregon.

Climate change, along with exotic species, disturbances, and land use change, will likely have major impacts on sagebrush steppe ecosystems in the western U.S. over the next century. To effectively manage sagebrush steppe landscapes for long-term goals, managers need information about the interacting impacts of climate change, disturbances and land management on vegetation condition. Using a climate-informed state-and-transition model, we evaluated the potential impacts of climate change on rangeland condition in central Oregon and the effectiveness of multiple management strategies. Under three scenarios of climate change, we projected widespread shifts in potential vegetation types over the twenty-first century, with declining sagebrush steppe and expanding salt desert shrub likely by the end of the century. Many extreme fire years occurred under all climate change scenarios, triggering rapid vegetation shifts. Increasing wildfire under climate change resulted in expansion of exotic grasses but also decreased juniper encroachment relative to projections without climate change. Restoration treatments in warm-dry sagebrush steppe were ineffective in containing exotic grass, but juniper treatments in cool-moist sagebrush steppe substantially reduced the rate of juniper encroachment, particularly when prioritized early in the century. Overall, climate-related shifts dominated future vegetation patterns, making management for improved rangeland condition more difficult. Our approach allows researchers and managers to examine long-term trends and uncertainty in rangeland vegetation condition and test the effectiveness of alternative management actions under projected climate change.

Restoration of mountain big sagebrush steppe following prescribed burning to control western juniper.

Western juniper (Juniperus occidentalis ssp. occidentalis Hook) encroachment into mountain big sagebrush (Artemisia tridentata spp. vaseyana (Rydb.) Beetle) steppe has reduced livestock forage production, increased erosion risk, and degraded sagebrush-associated wildlife habitat. Western juniper has been successfully controlled with partial cutting followed by prescribed burning the next fall, but the herbaceous understory and sagebrush may be slow to recover. We evaluated the effectiveness of seeding perennial herbaceous vegetation and sagebrush at five sites where juniper was controlled by partially cutting and prescribed burning. Treatments tested at each site included an unseeded control, herbaceous seed mix (aerially seeded), and the herbaceous seed mix plus sagebrush seed. In the third year post-treatment, perennial grass cover and density were twice as high in plots receiving the herbaceous seed mix compared to the control plots. Sagebrush cover and density in the sagebrush seeded plots were between 74- and 290-fold and 62- and 155-fold greater than the other treatments. By the third year after treatment, sagebrush cover was as high as 12 % in the sagebrush seeded plots and between 0 % and 0.4 % where it was not seeded. These results indicate that aerial seeding perennial herbaceous vegetation can accelerate the recovery of perennial grasses which likely stabilize the site. Our results also suggest that seeding mountain big sagebrush after prescribed burning encroaching juniper can rapidly recover sagebrush cover and density. In areas where sagebrush habitat is limited, seeding sagebrush after juniper control may increase sagebrush habitat and decrease the risks to sagebrush-associated species.

Critical phases in the seed development of common juniper (Juniperus communis).

Common juniper (Juniperus communis L.) populations in northwest European lowlands are currently declining in size and number. An important cause of this decline is a lack of natural regeneration. Low seed viability seems to be one of the main bottlenecks in this process. Previous research revealed a negative relation between seed viability and both temperature and nitrogen deposition. Additionally, the seeds of common juniper have a variable ripening time, which possibly influences seed viability. However, the underlying mechanisms remain unresolved. In order to elucidate this puzzle, it is important to understand in which phases of seed production the main defects are situated, together with the influence of ripening time. In this study, we compared seed viability of populations with and without successful recruitment. We examined three seed phases: (i) gamete development; (ii) fertilisation and early-embryo development; and (iii) late-embryo development. After the first two phases, we found no difference in the percentage viable seeds between populations with or without recruitment. After late-embryo development, populations without recruitment showed a significantly lower percentage of viable seeds. These results suggest that late-embryo development is a bottleneck in seed development. However, the complex interaction between seed viability and ripening time suggest that the causes should be in the second seed phase, as the accelerated development of male and female gametophytes may disturb the male-female synchrony for successful mating.