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Gene flow among different taxonomic units: evidence from nuclear and cytoplasmic markers in Cedrus plantation forests.

Hybridization and introgression are important natural evolutionary processes that can be successfully investigated using molecular markers and open- and controlled-pollinated progeny. In this study, we collected open-pollinated seeds from Cedrus atlantica, Cedrus libani and C. libani x C. atlantica hybrids from three French-plantation forests. We also used pollen from C. libani and Cedrus brevifolia to pollinate C. atlantica trees. The progeny were analyzed using three different types of molecular markers: RAPDs, AFLPs and cpSSRs. Chloroplast DNA was found to be paternally inherited in Cedrus from the progeny of controlled-crosses. Heteroplasmy, although possible, could not be undoubtedly detected. There was no indication of strong reproductive isolating barriers among the three Mediterranean Cedrus taxa. Gene flow between C. atlantica and C. libani accounted for 67 to 81% of viable open-pollinated seedlings in two plantation forests. We propose that Mediterranean Cedrus taxa should be considered as units of a single collective species comprising two regional groups, North Africa and the Middle East. We recommend the use of cpSSRs for monitoring gene flow between taxa in plantation forests, especially in areas where garden specimens of one species are planted in the vicinity of selected seed-stands and gene-conservation reserves of another species.

Cedrus↗

Abietoid seed fatty acid compositions--a review of the genera Abies, Cedrus, Hesperopeuce, Keteleeria, Pseudolarix, and Tsuga and preliminary inferences on the taxonomy of Pinaceae.

The seed fatty acid (FA) compositions of Abietoids (Abies, Cedrus, Hesperopeuce, Keteleeria, Pseudolarix, and Tsuga) are reviewed in the present study in conclusion to our survey of Pinaceae seed FA compositions. Many unpublished data are given. Abietoids and Pinoids (Pinus, Larix, Picea, and Pseudotsuga)-constituting the family Pinaceae-are united by the presence of several delta5-olefinic acids, taxoleic (5,9-18:2), pinolenic (5,9,12-18:3), coniferonic (5,9,12,15-1 8:4), keteleeronic (5,11-20:2), and sciadonic (5,11,14-20:3) acids, and of 14-methyl hexadecanoic (anteiso-17:0) acid. These acids seldom occur in angiosperm seeds. The proportions of individual delta5-olefinic acids, however, differ between Pinoids and Abietoids. In the first group, pinolenic acid is much greater than taxoleic acid, whereas in the second group, pinolenic acid is greater than or equal to taxoleic acid. Moreover, taxoleic acid in Abietoids is much greater than taxoleic acid in Pinoids, an apparent limit between the two subfamilies being about 4.5% of that acid relative to total FA. Tsuga spp. appear to be a major exception, as their seed FA compositions are much like those of species from the Pinoid group. In this respect, Hesperopeuce mertensiana, also known as Tsuga mertensiana, has little in common with Abietoids and fits the general FA pattern of Pinoids well. Tsuga spp. and H. mertensiana, from their seed FA compositions, should perhaps be separated from the Abietoid group and their taxonomic position revised. It is suggested that a "Tsugoid" subfamily be created, with seed FA in compliance with the Pinoid pattern and other botanical and immunological criteria of the Abietoid type. All Pinaceae genera, with the exception of Pinus, are quite homogeneous when considering their overall seed FA compositions, including delta5-olefinic acids. In all cases but one (Pinus), variations from one species to another inside a given genus are of small amplitude. Pinus spp., on the other hand, have highly variable levels of delta5-olefinic acids in their FA compositions, particularly when sections (e.g., Cembroides vs. Pinus sections) or subsections (e.g., Flexiles and Cembrae subsections from the section Strobus) are compared, although they show qualitatively the same FA patterns characteristic of Pinoids. Multicomponent analysis of Abietoid seed FA allowed grouping of individual species into genera that coincide with the same genera otherwise characterized by more classical botanical criteria. Our studies exemplify how seed FA compositions, particularly owing to the presence of delta5-olefinic acids, may be useful in sustaining and adding some precision to existing taxonomy of the major family of gymnosperms, Pinaceae.

Abies↗

Local NO3- or NH4+ supply modifies the root system architecture of Cedrus atlantica seedlings grown in a split-root device.

To study the effects of local nitrate or ammonium supply on the architecture of the Cedrus atlantica root system, cedar seedlings were grown in split-root boxes in a growth chamber. In each box-compartment, roots were fertilized with a solution containing nitrogen, either as nitrate [Ca(NO(3))(2)] or ammonium (NH(4)Cl), supplied at 0.1 or 5.0mM. For each seedling, the shoot growth was measured twice a week for 3 months. The root system architecture was also recorded twice a week by tracing the root elongation through the transparent face of the root observation boxes. The apical diameter of the tap-root relay and that of a representative sample of lateral roots were recorded once a month using a monocular magnifier. The increase of ammonium or nitrate concentration in the nutrient solution has significantly enhanced the production of lateral roots on the tap-root relay. After 90 days of culture, percentages of short lateral roots obtained with nitrate were higher than those obtained using ammonium. A preferential carbon allocation to the shoots was also obtained with an increasing nitrogen supply. Until the 40th day of culture, the elongation of lateral roots was similar for all treatments and ranged from 0.25 to 0.5 cm day(-1). From the 40th day to the 95th day, significant differences were observed between the compared modes and maximum elongation rates were obtained with 5mM NH(4)(+) (2.18 cm day(-1)) and 5mM NO(3)(-) (1.18 cm day(-1)). Local applications of nitrate and ammonium at a low or a high concentration had local effects on elongation and branching of the root system in the fertilized compartment. Contrasting effects of ammonium and nitrate were observed on the apical diameter of tap-roots and lateral roots. The root-split culture device confirmed that nitrate had local effects on the architecture of the C. atlantica root system.

Biomass↗

A novel lignan composition from Cedrus deodara induces apoptosis and early nitric oxide generation in human leukemia Molt-4 and HL-60 cells.

AP9-cd, a standardized lignan composition from Cedrus deodara consisting of (-)-wikstromal, (-)-matairesinol, and dibenzyl butyrolactol, showed cytotoxicity in several human cancer cell lines reported earlier. An attempt was made in this study to investigate the mechanism of cell death in human leukemia Molt-4 and HL-60 cells. It inhibited Molt-4 cell proliferation with 48-h IC(50) of approximately 15 microg/ml, increased sub-G0 cell fraction with no mitotic block, produced apoptotic bodies and induced DNA ladder formation. Flow cytometric analysis of annexinV-FITC/PI-stained cells showed time-related increase in apoptosis and post-apoptotic necrosis. All these biological end-points indicated cell death by apoptosis. Further, initial events involved massive nitric oxide (NO) formation within 4 h with subsequent late appearance of peroxides in cells; measured by flow cytometry using specific fluorescent probes. Persistently high levels of NO and peroxide appeared to decrease mitochondrial membrane potential (Psi(mt)) which was recovered by cyclosporin A in Molt-4 cells. AP9-cd caused 2-fold activation of caspase-3 in Molt-4 and 5-fold activation in HL-60 cells. Also caspases-8 and -9 were activated in HL-60 cells. Ascorbate suppressed the enhanced caspases activities indicating a pro-oxidant effect of AP9-cd. Further, caspase-3 activation correlated with NO generation that was partially impaired by nitric oxide synthase (NOS) inhibitors and ascorbate suggesting a role of pro-oxidant species in caspase-3 activation. AP9-cd produced no cytotoxicity in primary rat hepatocyte culture at the concentrations used. The studies indicated that AP9-cd mediated early NO formation leads to caspases activation, peroxide generation, and mitochondrial depolarization which may be responsible for mitochondrial-dependent and -independent apoptotic pathways involved in the killing of leukemia cells by AP9-cd.

Animals↗

Abietane diterpenes from the cones of Cedrus atlantica.

Five new abietanes, three of them isolated as the corresponding acetate derivatives, i.e., 9alpha,13alpha-epidioxiabiet-8(14)-en-18-ol, 7alpha,18-diacetoxy, 9beta,13beta-epidioxiabiet-8(14)-ene, 7alpha,18-diacetoxyabiet-8(14)-en-13beta-ol, 7alpha,18-diacetoxy-13beta-methoxyabiet-8(14)-ene, and 13beta-hydroxyabiet-8(14)-en-7-one, were isolated from the neutral part of the hexane extract of the cones of Cedrus atlantica collected in Middle Atlas, Morocco. The structures of these compounds were established by spectroscopic techniques, including 2D NMR spectra, and in the case of 1, by chemical correlation. The cytotoxicity of these abietane diterpenoids was tested against five cell lines.

Abietanes↗

Combinations of Azadirachta indica and Cedrus deodara oil with piperonyl butoxide, MGK-264 and Embelia ribes against Lymnaea acuminata.

The binary and tertiary combinations of plant-derived molluscicides Azadirachta indica and Cedrus deodara oil with synergists MGK-264, piperonyl butoxide (PB) and fruit powder of Embelia ribes were used against the Lymnaea acuminata. It was observed that the toxic effects of these mixtures were time- and dose-dependent. The binary and tertiary mixtures of plant-derived molluscicides with synergists were more toxic with respect to the single treatment of the plant-derived molluscicides. Maximum synergistic action in binary and tertiary combinations was found in A. indica + C. deodara oil and A. indica+ PB + C. deodara in 1:7 and 1:5:7 ratio, respectively.

Animals↗

Free radical scavenging active components from Cedrus deodara.

An activity-directed fractionation and purification process was used to identify the antioxidant components of Cedrus deodara. Dried heartwood powder of C. deodara was first defatted with petroleum ether and then extracted with chloroform. The chloroform extract showed strong antioxidant activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. This fraction was then subjected to separation and purification using silica gel column chromatography. Three compounds with potent antioxidant activity were isolated in significant yields and identified by spectroscopic methods ((1)H NMR, (13)C NMR, IR, and MS). They were identified as (-)-matairesinol, (-)-nortrachelogenin, and a dibenzylbutyrolactollignan (4,4',9-trihydroxy-3,3'-dimethoxy-9,9'-epoxylignan). This is the first report of the occurrence of these compounds in C. deodara.

Antioxidants↗

Antibacterial diterpenoids from Cedrus atlantica.

Four diterpene alcohols were isolated from the neutral hexane extract of the cones of Cedrus atlantica, and their structures were confirmed after comparing their spectral data with literature values. These products exhibited significant antibacterial activity against gram (+/-) bacteria.

Alcohols↗

Pollen production, microsporangium dehiscence and pollen flow in Himalayan cedar (Cedrus deodara Roxb. ex d. Don).

Microsporangium dehiscence, pollen production and dispersal were studied in Himalayan cedar (Cedrus deodara) during 1998 and 1999. Microsporangium dehiscence showed diurnal periodicity and was found to be related to air temperature and relative air humidity, with a strobilus taking 2 d to dehisce completely in warmer conditions and 3 d in cooler ones. The frequency of flowering in C. deodara was highly variable during the two successive years; however, cyclical production of pollen grains was observed in 50% of the trees. The maximum concentration of pollen grains in the air was found between 1200 and 1600 h, and this period was also noted to be the best time for pollination. Studying migration of pollen grains from isolated single trees in three directions showed that migration was not uniform in all directions. Long-distance transport of pollen grains was observed in the downhill direction. However, in the uphill and horizontal directions grains could travel only up to 97.5 and l95.1 m, respectively, and the frequency of pollen grains to the source frequency at these distances was only 1.9 and 2-5%, respectively. The results suggest that an isolation barrier of 190 m may be considered as a minimum for the management of deodar seed orchards.

Air↗

Root system architecture and receptivity to mycorrhizal infection in seedlings of Cedrus atlantica as affected by nitrogen source and concentration.

Effects of nitrogen (N) source and concentration on root system architecture and receptivity to mycorrhizal infection were studied in seedlings of Atlas cedar (Cedrus atlantica Manetti) grown in root observation boxes in a controlled-environment chamber. Nitrogen was supplied in a solution containing either NO3-; or NH4+ at a concentration of either 0.25 or 5.0 mM. Root extension was recorded twice weekly by tracing the roots growing in contact with the transparent face of the root observation box. Among treatments, lateral root production and branching density were greatest with 5.0 mM NO3-. Inoculation with mycelium of Tricholoma cedrorum Malencon was carried out 3 months after the start of the N treatments. The highest percentage of mycorrhizal roots, and the greatest amounts of living mycelium (estimated by the ergosterol assay) were observed in the NO3- treatments. Differences in root branching density among the N treatments were insufficient to explain the observed differences among treatments in the extent of mycorrhizal infection of seedlings.

Agaricales↗

Evaluation of some competition indices for the construction of Lebanon cedar (Cedrus libani A. Rich.) stand models.

The purpose of this research is to test the precision of some published competition indices of Lebanon cedar (Cedrus libani A. Rich.) for the estimation of future periodic diameter increment of individual trees. Twenty- nine published competition indices were tested, using fifteen separate sets of data and their pooled values, collected from various stand age and site quality classes Lebanon cedar at Antalya. Temporary sample plots were taken in Elmali-Qamkuyusu (9 sample plots) and Finike-Pinarcik (6 sample plots) in 2001. Every plot was stem mapped (x and y coordinate system), diameter (dbh), total height, crown length, crown diameter and 10-year radial increment were recorded for trees greater than 4 cm in dbh. Then, in order to evaluate these competition indices for the prediction of the periodic diameter increment of the individual trees. Three linear models have been constructed for each competition index. It was found that the competition indices (Daniels et al., 1986; Biging and Dobbertin, 1995; Pukkala and Kolström, 1987; Hegyi, 1974) with larger influence-zone areas produce better results.

Cedrus↗

Spasmolytic constituents of Cedrus deodara (Roxb.) Loud: pharmacological evaluation of himachalol.

Himachalol has been identified as the major antispasmodic constituent in the wood of Cedrus deodara. The pharmacological studies of himachalol on various isolated smooth muscles (guinea pig ileum, rabbit jejunum, rat uterus, and guinea pig seminal vesicle) and against different agonists (acetylcholine, histamine, serotonin, nicotine, and barium chloride) indicated spasmolytic activity similar to that of papaverine. It was a more potent antagonist of barium chloride-induced spasm of guinea pig ileum than papaverine but less effective in reverting a similar spasm of rabbit jejunum and had no relaxing effect alone. In the conscious immobilized cat, intragastric administration of himachalol or papaverine (100 mg/kg) produced equal inhibition of carbachol-induced spasm of the intestine, lasting about 2 hr, but himachalol had a faster onset of action. Himachalol was devoid of spasmolytic effect on the bronchial musculature of guinea pig but was 3.3 times more potent than papaverine in antagonizing epinephrine-induced contraction of the guinea pig seminal vesicle. Intravenous injection of himachalol (3-10 mg/kg) in the cat produced a dose-dependent fall in blood pressure and an increased femoral blood flow.

Animals↗

Atmospheric PCDD/F and PCB levels implicated by pine (Cedrus deodara) needles at Dalian, China.

Dalian is a seaside city situated in the northeastern monsoon area of China. For the first time, levels of PCDD/F and PCB congeners in pine (Cedrus deodara) needles of Dalian urban areas were investigated. Two sampling campaigns with 17 sampling points were performed in 2002. The summation of tetra- to octachlorinated PCDD/Fs and summation of 209 PCB congeners in Dalian pine needles averaged 127+/-40 ng/kg (dry) and 4389+/-1575 ng/kg (dry), respectively. Average toxic equivalence (TEQ) for PCDD/Fs and PCBs are 2.1 and 0.4 ng/kg (dry), respectively. The pine needles can differentiate spatial variation of the pollutants. The PCDD/F and PCB levels in Dalian pine needles are low or comparable with other international regions that were not impacted by evident point sources. The data can serve as a base for long-term spatial and temporal studies of PCDD/Fs and PCBs in China.

China↗

Studies on the anti-inflammatory and analgesic activity of Cedrus deodara (Roxb.) Loud. wood oil.

The volatile oil extracted by steam distillation of the wood of Cedrus deodara was examined for its oral anti-inflammatory and analgesic activity at the doses of 50 and 100 mg/kg body weight. It produced significant inhibition of carrageenan-induced rat paw edema and of both exudative-proliferative and chronic phases of inflammation in adjuvant arthritic rats at doses of 50 and 100 mg/kg body weight. The oil at both tested doses was found to possess analgesic activity against acetic acid-induced writhing and hot plate reaction in mice.

Acetic Acid↗

Clinical and immunologic evaluation of Cedrus deodara pollen: a new allergen from India.

BACKGROUND: Allergy to pollen from gymnosperms is well documented in the West. However, many allergenic species are native to the Himalayan region of India, and Cedrus deodara (Pinaceae) was selected for allergologic investigation. The objective was to define the allergologic and immunochemical aspects of C. deodara pollen. METHODS: Pollen antigen from C. deodara (CD) was prepared and characterized by biochemical and biologic assays. Specific IgE binding was determined by means of ELISA and immunoblotting. RESULTS: CD pollen antigen caused marked skin sensitivity in 7.5% of an atopic population. A significantly elevated level of CD-specific IgE antibodies was observed in 65.8% of the skin-positive patients. Immunoblotting showed protein fractions of 37, 44, 58, and 78 kDa with 100% binding with the patients' sera suspected to be due to carbohydrate moieties. CONCLUSIONS: Patients from the Himalayan region, where CD occurs naturally, were sensitized more than patients from distant places. The immunochemical characterization revealed multiple protein fractions from low to very high molecular mass (14-126 kDa) mostly in the acidic pI range. CD pollen has been recognized as a new allergen from India for the first time. The role of pollen as a causative agent of respiratory allergic disorders is very well established, as is evident from the recent increase of reports from across the world (1-4). India is blessed with the richest flora on the earth, from alpine tundra to Rajasthan desert. Consequently, it provides considerable variation in the quality and quantity of airborne pollen in different ecogeographic regions of the country (5-8). Although studies on the allergenic properties of airborne pollen from various species have been carried out by several workers in India (9-12), information on allergy to aerial pollen from Himalayan tree species has been completely

Allergens↗

Evidence of variation in segregation patterns within a Cedrus population.

We used horizontal starch-gel electrophoresis for a genetic analysis of isozymes within one French Cedrus atlantica stand. Eleven to 29 megagametophytes per tree from 186 trees were assayed. Among the 33 enzyme systems tested, 15 correctly resolved and 8 appeared variable in at least one zone of activity: ACP, GOT, IDH, LAP, MDH, MNR, PGI, and SKDH. They were coded by at least 12 polymorphic loci which were described and tested for Mendelian segregation and linkage. Segregation patterns and linkage relationships were variable in the population. We detected homogeneous segregation distortion for loci Idh, Acp-c, and Got-a over the whole set of segregating progeny. We also found segregation distortions in a significant proportion of progeny for loci Got-b, Mdh-c, Pgi-b, and SKDH: The Acp-c and Got-b loci were linked with an overall map distance of 17 cM, but distance varied drastically among progeny. Both segregation distortions and heterogeneity of recombination frequencies indicate the occurrence of a genetic load in this population.

Chromosome Segregation↗

Antimicrobial activities of several parts of Pinus brutia, Juniperus oxycedrus, Abies cilicia, Cedrus libani and Pinus nigra.

In this study, the antimicrobial activities of several parts of various trees grown in the Kahramanmaraş region of Turkey were investigated by the disc diffusion method. Chloroform, acetone and methanol extracts of leaves, resins, barks, cones and fruits of Pinus brutia Ten., Juniperus oxycedrus L., Abies cilicia Ant. & Kotschy Carr., Cedrus libani A. Rich. and Pinus nigra Arn. were prepared and tested against Bacillus megaterium DSM 32, Bacillus subtilis IMG 22, Bacillus cereus FMC 19, Escherichia coli DM, Klebsiella pneumoniae FMC 3, Enterobacter aerogenes CCM 2531, Staphylococcus aureus Cowan 1, Mycobacterium smegmatis RUT, Proteus vulgaris FMC 1, Listeria monocytogenes Scoot A, Pseudomonas aeruginosa DSM 5007, Candida albicans CCM 314, Candida tropicalis MDC 86 and Penicillium italicum K. The results showed that antifungal effects were not observed for the whole extracts, E. coli was not inhibited by any of the plant extracts except by the chloroform and acetone extracts of the leaves of A. cilicia, which showed inhibition zones of 16-18 mm, respectively. All the plant extracts used in this study inhibited the development of the other bacteria studied. When the results of this study were compared with an ampicillin standard, it was found that the microorganisms studied were generally susceptible, intermediate or resistant to the extracts of species when compared with the ampicillin standard. On the other hand, the acetone and methanol extracts of Juniperus fruits were found to be quite resistant.

Anti-Bacterial Agents↗