Effect of Sclerotinia sclerotiorum on the disease development,growth, oil yield and biochemical changes in plants of Mentha arvensis

Experiment was carried out to determine the effect of Sclerotinia sclerotiorum on the disease development, growth, oil yield and biochemical changes in the plants of Mentha arvensis. With the increase in initial inoculum levels of S. sclerotiorum a corresponding decrease in plant fresh and dry weights were recorded. The maximum reduction in the shoot-roots/suckers fresh weight and shoot-roots/suckers dry weights (39.8%, 43.6%, 40.3% and 42.9%), respectively, was observed at the highest initial inoculum level of 12 g fungal mycelium/5 kg soil as compared to uninoculated control. The infection of roots and suckers due to S. sclerotiorum increased with increasing initial inoculum levels. At the lowest initial inoculum (1.0 g mycelium/5 kg soil), infection was observed 18.0% and at the highest (12 g mycelium/5 kg soil), it was 80.2%. Significant (P ⩽ 0.01) reduction in oil yield, total chlorophyll, total phenol and total sugar content of M. arvensis plants was observed at the lowest inoculum level as compared to uninoculated control.     

Optimization of phytase production by Penicillium purpurogenum GE1 under solid state fermentation by using Box–Behnken design

Phytase production by Penicillium purpurogenum GE1 isolated from soil around bean root nodules was investigated by solid state fermentation (SSF) using mixed substrates consisted of corn cob and corn bran. The SSF conditions were optimized by using one-variable–at-a-time strategy. The optimum conditions for phytase production were at 27 °C, pH 8 and 66% moisture content. The study of different carbon and nitrogen sources revealed that glucose and peptone registered the highest enzyme productivity (92 ± 5.6 U/g ds, 125 ± 4.9 U/g ds). Among different surfactants, maximum phytase productivity was observed with Tween 80 at 0.001 concentrations (170 ± 4.2 U/g ds). A Box–Behnken design was employed to investigate the optimization of the most significant variables affecting the enzyme production. Maximal phytase production was detected after the addition of (g/5 g ds): 0.75 glucose, 0.375 peptone and 0, 01 tween 80. This result represented an improvement in phytase production of 2.6 folds when compared to that previously obtained using the basal medium under the same cultivation conditions. The generated model was found to be very adequate for phytase production (90% accuracy) as the experimental value was 444 ± 3.5 U/g ds compared to 401 U/g ds for the predicted value. In brief, the production of phytase using corn cob and corn bran is a novel and cheap way for the production of this important enzyme and opens a new way for researchers to discover and explore this arena.     

Thermus parvatiensis RLT sp. nov., Isolated from a Hot Water Spring,Located Atop the Himalayan Ranges at Manikaran,India

A Gram negative, yellow pigmented, rod shaped bacterium designated as RL^T was isolated from a hot water spring (90–98 °C) located at Manikaran in Northern India. The isolate grows at 60–80 °C (optimum, 70 °C) and at pH 7.0–9.0 (optimum pH 7.2). Phylogenetic analysis of 16S rRNA gene sequences and levels of DNA–DNA relatedness together indicate that the new isolate represents a novel species of the genus Thermus with closest affinity to Thermus thermophilus HB8^T (99.5 %) followed by Thermus arciformis (96.4 %). A comparative analysis of partial sequences of housekeeping genes (HKG) further revealed that strain RL^T is a novel species belonging to the genus Thermus. The melting G+C content of strain RL^T was calculated as 68.7 mol%. The DNA–DNA relatedness value of strain RL^T with its nearest neighbours (>97 %) was found to be less than 70 % indicating that strain RL^T represents a novel species of the genus Thermus. MK-8 was the predominant respiratory quinone. The presence of characteristic phospholipid and glycolipid further confirmed that strain RL^T belongs to the genus Thermus. The predominant fatty acids of strain RL^T were iso-C_17:0 (23.67 %) and iso-C_15:0 (24.50 %). The results obtained after DNA–DNA hybridization, biochemical and physiological tests clearly distinguished strain RL^T from its closely related species. Thus, strain RL^T represents a novel species of the genus Thermus for which the name Thermus parvatiensis is proposed (=DSM 21745^T= MTCC 8932^T).

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0538-4) contains supplementary material, which is available to authorized users.     

A new species of plasmodiidae (Coccidia: Hemosporidia) from the blood of the skink Scincus hemprichii (Scincidae: Reptilia) in Saudi Arabia

Fallisia arabica n. sp. was described from peripheral blood smears of the Skink lizard, Scincus hemprichii from Jazan Province in the southwest of Saudi Arabia. Schizogony and gametogony take place within neutrophils in the peripheral blood of the host. Mature schizont is rosette shaped 17.5 ± 4.1 × 17.0 ± 3.9 μm, with a L/W ratio of 1.03(1.02–1.05) μm and produces 24(18–26) merozoites. Young gametocytes are ellipsoidal, 5.5 ± 0.8 × 3.6 ± 0.5 μm, with a L/W of 1.53(1.44–1.61) μm. Mature macrogametocytes are ellipsoidal, 9.7 ± 1.2 × 7.8 ± 1.0 μm, with a L/W of 1.24(1.21–1.34) μm and microgametocytes are ellipsoidal, 7.0 ± 1.1 × 6.8 ± 0.9 μm. with a L/W of 1.03(1.01–1.10) μm. In comparison to the described Fallisia species, this new taxon has rosette schizonts and is larger than F. dominicensis, in Hispaniola, F. bipocrati, F. poecilopi, in Panama, F. thecadactyli in Venezuela, and F. effusa, F. simplex, F. modesta, in Brazil. F. arabica has fewer merozoites than F. effusa, F. poecilopi, F. thecadactyli and F. siamense in Thailand. This new species has more merozoites than F. dominicensis and F. modesta. All of these species belong to diverse saurian families (Agamidae, Gekkonidae, Polychrotidae, Scincidae and Teiidae) parasitize only thrombocytes or lymphocytes and some species parasitize immature erythroid cells and leucocytes.     

De-hairing protease production by an isolated Bacillus cereus strain AT under solid-state fermentation using cow dung: Biosynthesis and properties

Agro-industrial residues and cow dung were used as the substrate for the production of alkaline protease by Bacillus cereus strain AT. The bacterial strain Bacillus cereus strain AT produced a high level of protease using cow dung substrate (4813 ± 62 U g⁻¹). Physiological fermentation factors such as the incubation time (72 h), the pH (9), the moisture content (120%), and the inoculum level (6%) played a vital role in the enzyme bioprocess. The enzyme production improved with the supplementation of maltose and yeast extract as carbon and nitrogen sources, respectively. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and zymogram analysis of the purified protease indicated an estimated molecular mass of 46 kDa. The protease enzyme was stable over a temperature range of 40–50 °C and pH 6–9, with maximum activity at 50 °C and pH 8. Among the divalent ions tested, Ca²⁺, Na⁺ and Mg²⁺ showed activities of 107 ± 0.7%, 103.5 ± 1.3%, and 104.6 ± 0.9, respectively. The enzyme showed stability in the presence of surfactants such as sodium dodecyl sulfate and on various commercially available detergents. The crude enzyme effectively de-haired goat hides within 18 h of incubation at 30 °C. The enzymatic properties of this protease suggest its suitable application as an additive in detergent formulation and also in leather processing. Based on the laboratory results, the use of cow dung for producing and extracting enzyme is not cumbersome and is easy to scale up. Considering its cheap cost and availability, cow dung is an ideal substrate for enzyme bioprocess in an industrial point of view.     

Heterologous Expression and Efficient Secretion of Chitosanase from Microbacterium sp. in Escherichia coli

A recombinant expression vector, pCT7-CHISP6H, was constructed for the secretory expression of mature peptide of chitosanase (mMschito) from Microbacterium sp. OU01. The vector contains several elements, including T7 promoter, signal peptide sequence of mschito, 6 × His-tag sequence and PmaCI restriction enzyme cloning site. In pCT7-CHISP6H, mMschito was fused into signal peptide sequence of mschito gene to construct recombinant plasmid pCT7-CHISP6H-mMschito. The recombinant plasmid was transformed into Escherichia coli BL21(DE3) and then expressed. The recombinant protein was secreted into the Luria–Bertani broth and the chitosanase activity in supernatant of the culture could reach up to 67.56 U/mL. The rmMschito in the broth supernatant was purified using HisTrap™ FF Crude column and the purified rmMschito was shown to be apparent homogeneity by 12 % SDS–PAGE analysis. Detected by 4700 MALDI-TOF–TOF-MS, the molecular weight of the purified rmMschito was 26,758.1875 and it was consistent with the predicted molecular weight. Chitosan (degree of deacetylation of 99 %) was mostly hydrolyzed into chitopentaose, chitotriose, and chitobiose by the purified rmMschito.     

Three‐liquid‐phase salting‐out extraction of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)‐rich oils from Euphausia superba

The TLPSOES parameters were optimized by response surface methodology using Box–Behnken design, which were 16.5% w/w of ammonium citrate, 17.5% w/w of ethanol, and 46% w/w of n‐hexane at 70 min of stirring time. Under optimized conditions the extraction efficiency attained was 90.91 ± 0.97% of EPA, 90.02 ± 1.04% of DHA, and 91.85 ± 1.11% of KO in the top n‐hexane phase. The highest extraction efficiency of proteins and flavonoids, i.e. 88.34 ± 1.35% and 79.67 ± 1.13%, was recorded in the solid interface and ethanol phase, respectively. The KO extracted by TLPSOES system consisted of lowest fluoride level compared to the conventional method and whole wet krill biomass. The TLPSOES is a potential candidate for nutraceutical industry of KO extraction from wet krill biomass.     

Protection of human erythrocyte using Crinumasiaticum extract and lycorine from oxidative damage induced by 2-amidinopropane

The intention of this investigation was to evaluate the free radical scavenging activity and erythrocyte protective activity of ethanolic extract of Crinumasiaticum (L) and lycorine. The ethanolic extract of C. asiaticum (L) and lycorine were found to have different levels of antioxidant properties in the test models. Both ethanolic extract of C. asiaticum (L) (0.5–2.5 mg/ml) and lycorine (0.010 mg–0.050 mg/ml) increases the percentage of lipid peroxidation inhibition (26.25 ± 0.23% and 19.25 ± 0.23%) and enhances the free radical scavenging activity (20.92 ± 0.22% and 20.52 ± 0.22%), scavenging of hydrogen peroxide (25.67 ± 0.17% and 23.07 ± 0.3%) superoxide anion scavenging activity (27.69 ± 0.16% and 16.09 ± 0.7%) at concentration of 2.5 and 0.050 mg of C. asiaticum (L) and lycorine, respectively. But compared with tocopherol (P < 0.05) less activity was observed by C. asiaticum (L) and lycorine. The ethanolic extract of C. asiaticum (L) and lycorine were normalized to reduce the level of glutathione and also to sustain the status of protein in erythrocytes during the peroxyl radical [2,2-azobis (2-amidinopropane) dihydrochloride (AAPH)] induced oxidative damage in ex vivo model. The present results of the investigations demonstrated that protective nature of the C. asiaticum (L) and lycorine will be considered as a significant natural antioxidant source.     

Effects of Riyadh cement industry pollutions on some physiological and morphological factors of Datura innoxia Mill. plant

Cement factory emissions into air cause serious air pollution and affect the plant and animal life in the environment. Herein, we report the effects of cement industry emissions (O₃, SO₂ and NO₂) in air, as pollutants, at Riyadh City on Datura innoxia Mill. plant. Morphological characters including plant height, leaves area and number, fresh and dry weight of shoot and root systems of D. innoxia showed a significant reduction from their normal control plants as a response to exposure to pollutant emissions. Chlorophyll and carotenoid contents recorded reductions in values compared to control plant, and the lowest values of chlorophyll A, B, total chlorophyll, carotenoids and total pigments were 0.431, 0.169, 0.60, 0.343 and 0.943 mg/g respectively at a distance of 1–5 m from the cement factory in fruiting stage. These changes in values may be attributed to a probable deceleration of the biosynthetic process rather than degradation of pigments. Further D. innoxia showed a significant (P < 0.01) reduction in non-reducing and total sugars, protein and total lipid contents compared with the control plant. The root system recorded the lowest values of reducing sugars (0.350 mg/g f. wt.), non-reducing sugars (0.116 mg/g f. wt.), total sugars (0.466 mg/g f. wt.), protein content (0.931 mg/g f. wt.) and total lipids content (0.669 mg/g f. wt.) in fruiting stage at a distance of 1–5 m from the cement factory. The peroxidase activity of shoot and root systems of the studied plant was also significantly higher than those of control plant. Thus a highest value of (29.616 units/g f. wt.) peroxidase activity was recorded in vegetative stage of shoot system at a distance 1–5 m from the cement factory. Results of the study indicated that cement industry emission strongly influence the physiology and morphology of date palm D. innoxia which contribute date fruits, a staple food in the Arab world.     

Genetic transformation of Indian isolate of Lemna minor mediated by Agrobacterium tumefaciens and recovery of transgenic plants

Transgenic plants of an Indian isolate of Lemna minor have been developed for the first time using Agrobacterium tumefaciens and hard nodular cell masses ‘nodular calli’ developed on the BAP - pretreated daughter frond explants in B₅ medium containing sucrose (1.0 %) with 2,4-D (5.0 μM) and 2-iP (50.0 μM) or 2,4-D (50.0 μM) and TDZ (5.0 μM) under light conditions. These calli were co-cultured with A. tumefaciens strain EHA105 harboring a binary vector that contained genes for β-glucuronidase with intron and neomycin phosphortransferase. Transformed cells selected on kanamycin selection medium were regenerated into fronds whose transgenic nature was confirmed by histochemical assay for GUS activity, PCR analysis and Southern hybridization. The frequency of transformation obtained was 3.8 % and a period of 11–13 weeks was required from initiation of cultures from explants to fully grown transgenic fronds. The pretreatment of daughter fronds with BAP, use of non-ionic surfactant, presence of acetosyringone in co-cultivation medium, co-culture duration of 3 d and 16 h photoperiod during culture were found crucial for callus induction, frond regeneration and transformation of L. minor. This transformation system can be used for the production of pharmaceutically important protein and in bioremediation.     

Effect of ultraviolet radiation on chlorophyll,carotenoid, protein and proline contents of some annual desert plants

Investigation was carried out to find whether enhanced ultraviolet radiation influences the Malva parviflora L., Plantago major L., Rumex vesicarius L. and Sismbrium erysimoids Desf. of some annual desert plants. The seeds were grown in plastic pots equally filled with a pre-sieved normal sandy soil for 1 month. The planted pots from each species were randomly divided into equal groups (three groups). Plants of the first group exposed to white-light tubes (400–700 nm) 60 w and UV (365 nm) 8 w tubes. The second group was exposed to white-light tubes (400–700 nm) 60 w and UV (302 nm) 8 w tubes. The third group was exposed to white-light tubes (400–700 nm) 60 w and UV (254 nm) 8 w tubes, respectively, for six days. The results indicated that the chlorophyll contents were affected by enhanced UV radiation. The chlorophyll a, b, and total contents were decreased compared with the control values and reduced with the enhanced UV radiation, but the carotenoid was increased compared with the control and also reduced with the enhanced UV radiation. So, the contents of chlorophylls varied considerably. M. parviflora showed the highest constitutive levels of accumulated chlorophyll a, b, and total chlorophyll (0.463, 0.307 and 0.774 mg g⁻¹ f w) among the investigated plant species. P. major showed the lowest constitutive levels of the chloroplast pigments, 0.0036, 0.0038 and 0.0075 mg g⁻¹ f w for chlorophyll a, b, and total chlorophyll at UV-365 nm, respectively. The protein content was decreased significantly in both root and shoot systems compared with the control values but, it was increased with increasing wave lengths of UV-radiation of all tested plants. R. vesicarius showed the highest protein contents among the investigated plants; its content was 3.8 mg g⁻¹ f w at UV-365 nm in shoot system. On the other hand, decreasing ultraviolet wave length induced a highly significant increase in the level of proline in both root and shoot of all tested plants. From the results obtained, it is suggested that proline can protect cells against damage induced by ultraviolet radiation. Statistically, the variations of the studied metabolic activities were significant due to UV radiation treatment in shoot and root system of all investigated plant species.     

Efficacy of leaves extract of Calotropis procera Ait. (Asclepiadaceae) in controlling Anopheles arabiensis and Culex quinquefasciatus mosquitoes

The present study aimed to investigate, the larvicidal, adult emergence inhibition and oviposition deterrent activity of aqueous leaves extract of Calotropis procera against Anopheles arabiensis and Culex quinquefasciatus as natural mosquito larvicide. The larvicidal activity was monitored against 2nd, 3rd and 4th instar larvae of each mosquito species 24 h post-treatment. Adult emergence inhibition activity was tested by exposing 3rd instar larvae of each mosquito species to different concentrations of extracts (200, 400, 600, 800 and 1000 ppm for An. arabiensis and 100, 200, 300, 400, 500 and 600 ppm for Cx. quinquefasciatus). Probit analysis was used to analyze data from bioassay experiments. The oviposition deterrent activity was tested by using three different concentrations of extracts (1000, 500 and 200 for An. arabiensis, and 1000, 500 and 100 for Cx. quinquefasciatus) that caused high, moderate and low larval mortality in the larvicidal experiment against 3rd instar larvae. It was found that, LC50–LC90 values calculated were 273.53–783.43, 366.44–1018.59 and 454.99–1224.62 ppm for 2nd, 3rd and 4th larval instars, respectively, of An. arabiensis and 187.93–433.51, 218.27–538.27 and 264.85–769.13 ppm for 2nd, 3rd and 4th larval instars, respectively, of Cx. quinquefasciatus. Fifty percent of adult emergence inhibition (EI50) was shown at 277.90 and 183.65 ppm for An. arabiensis and Cx. quinquefasciatus, respectively. The pupal stage was not affected till a concentration of 5000 ppm. The extract showed oviposition deterrence and effective repellence against both mosquito species at different concentrations, with the observation on that maximal eggs were laid in low concentration of extract. These results suggest that the leaves extract of C. procera possess remarkable larvicidal, adult emergence inhibitor, repellent and oviposition deterrent effect against both An. arabiensis and Cx. quinquefasciatus, and might be used as natural biocides for mosquito control.     

Genotype patterns at CLU,CR1, PICALM and APOE,cognition and Mediterranean diet: the PREDIMED-NAVARRA trial

The traditional Mediterranean diet (MedDiet) has shown beneficial effects on cognitive decline. Nevertheless, diet–gene interactions have been poorly evaluated. We aimed to investigate diet–gene interaction in the PREDIMED-NAVARRA randomized trial. A total of 522 participants (67 ± 6 years at baseline) enrolled in the PREDIMED-NAVARRA trial were randomly allocated to one of three diets: two MedDiets (supplemented with either extra-virgin olive oil or nuts) or a low-fat diet. They were evaluated with the Mini-Mental State Examination (MMSE) and the Clock Drawing Test (CDT) after 6.5 years of intervention. Subjects were genotyped for CR1-rs3818361, CLU-rs11136000, PICALM-rs3851179 and Apolipoprotein E (ApoE) genes. We studied MedDiet–gene interactions for cognition and assessed the effect of the MedDiet on cognition across different genetic profiles. A significant interaction (p = 0.041) between CLU-rs11136000 and the MedDiet intervention on the MMSE was found with a beneficial effect of MedDiet among carriers of the T minor allele (B = 0.97, 95 % CI 0.45–1.49). Similar effect was observed for CR1-rs3818361, but no significant interaction was observed (p = 0.335). For PICALM-rs3851179, the MedDiet intervention showed a beneficial effect in both genotype groups. No apparent interaction was found for the CDT between intervention and gene variants. Similarly, participants randomly allocated to MedDiet groups, with favorable profiles of CR1, CLU and PICALM genes, significantly improved CDT scores compared to controls with the same genetic profile. Cognitive performance was better for non-ApoE4 and for ApoE4 carriers of MedDiet groups compared to controls, but for CDT performance, we only found statistical significant differences for non-ApoE4 carriers. A MedDiet intervention modulates the effect of genetic factors on cognition. The effect of MedDiet might be greater for subjects with a more favorable genetic profile.     

Continuous and Batch Production of Chloroperoxidase by Mycelial Pellets of Caldariomyces fumago in an Airlift Fermentor

Batch and continuous production of the extracellular heme glycoprotein chloroperoxidase (CPO) was studied with an airlift fermentor. We induced Caldariomyces fumago CMI 89362 to form pellets by transferring a small inoculum volume in preculture prior to growth in a 1-liter fermentor. Continuous replacement of the fructose-salts medium (dilution rate, 0.008 h⁻¹) supported continuous CPO formation at an average concentration of 128 ± 10 mg of CPO liter⁻¹ for 8 days. Optimum CPO production rates averaged 1.2 ± 0.1 mg of CPO h⁻¹ at dilution rates below 0.033 h⁻¹. Varying the carbohydrate content of the feed solution or the time of starting the feed did not significantly alter the amount of CPO produced. Batch fermentation in the airlift fermentor resulted in maximum CPO concentrations of 280 ± 80 mg of CPO liter⁻¹, although on two separate occasions CPO concentrations reached 400 to 450 mg liter⁻¹, which was double the amount obtained by free hyphae in shake flask culture.     

Comparative Response of Alfalfa to Pratylenchus penetrans Populations

Four populations of Pratylenchus penetrans did not differ (P > 0.05) in their virulence or reproductive capability on Lahontan alfalfa. There was a negative relationship (r = -0 .7 9 ) between plant survival and nematode inocula densities at 26 ± 3 C in the greenhouse. All plants survived at an inoculum level (Pi) of 1 nematode/cm³ soil, whereas survival rates were 50 to 55% at 20 nematodes/cm³ soil. Alfalfa shoot and root weights were negatively correlated (r = - 0.87; P < 0.05) with nematode inoculum densities. Plant shoot weight reductions ranged from 13 % at Pi 1 nematode/cm³ soil to 69% for Pi 20 nematodes/cm³ soil, whereas root weight reductions ranged from 17% for Pi 1 nematode/cm³ soil to 75% for Pi 20 nematodes/cm³ soil. Maximum and minimum nematode reproduction (Pf/Pi) for the P. penetrans populations were 26.7 and 6.2 for Pi 1 and 20 nematodes/cm³ soil, respectively. There were negative correlations between nematode inoculum densities and plant survival (r = 0.84), and soil temperature and plant survival (r = -0 .7 8 ). Nematode reproduction was positively correlated to root weight (r = 0.89).     

Three new species of the genus Aporcelaimoides Heyns, 1965 from Vietnam (Nematoda,Dorylaimida, Aporcelaimidae), with an updated taxonomy of the genus

Three new species of Aporcelaimoides from natural habitats in Vietnam are studied, described and illustrated, including line drawings, LM and/or SEM pictures. Aporcelaimoides brevistylum sp. n. is characterized by its body 1.95–2.90 mm long, lip region offset by deep constriction and 17–18 µm broad, ventral side of mural odontostyle 11–14 µm long with aperture occupying 62–71% of its length, neck 663–767 µm long, pharyngeal expansion occupying 58–66% of total neck length, uterus a simple tube 85–182 µm long, pars refringens vaginae absent, V = 55–63, tail short and rounded (34–46 µm, c = 49–76, c’ = 0.6–0.8), spicules 67–86 µm long, and one ventromedian supplement out the range of spicules. Aporcelaimoides minor sp. n. is distinguished in having body 2.09–2.61 mm long, lip region offset by deep constriction and 19–20 µm broad, mural odontostyle 14–16 µm long at its ventral side with aperture occupying 73–84% of its length, neck 579–649 µm long, pharyngeal expansion occupying 57–66% of total neck length, uterus a simple tube 44–69 µm long, pars refringens vaginae well developed, V = 48–56, female tail very short, rounded conoid or truncate (14–26 µm, c = 90–146, c’ = 0.3–0.6), and male unknown. Aporcelaimoides silvaticum sp. n. is characterized by its body 2.09–2.60 mm long, lip region offset by depression and 17–18 µm broad, mural odontostyle 11–12 µm long at its ventral side with aperture occupying 60–66% of its length, neck 597–720 µm long, pharyngeal expansion occupying 58–64% of total neck length, uterus a simple tube 128–243 µm long, pars refringens vaginae well developed, V = 58–60, tail short and rounded (27–37 µm, c = 67–94, c’ = 0.6–0.7), spicules 64–75 µm long, and two or three widely spaced ventromedian supplements bearing hiatus. The genus Aporcelaimoides is restored, its diagnosis emended, and three species of Sectonema, namely Sectonema amazonicum, Sectonema haguei and Sectonema moderatum, transferred to it. An updated list of its species, a key to their identification and a tabular compendium with the most important morphometric features are also presented.     

Interactive effects of elevated CO2 and precipitation change on leaf nitrogen of dominant Stipa L. species

Nitrogen (N) serves as an important mineral element affecting plant productivity and nutritional quality. However, few studies have addressed the interactive effects of elevated CO₂ and precipitation change on leaf N of dominant grassland genera such as Stipa L. This has restricted our understanding of the responses of grassland to climate change. We simulated the interactive effects of elevated CO₂ concentration and varied precipitation on leaf N concentration (N_mass) of four Stipa species (Stipa baicalensis, Stipa bungeana, Stipa grandis, and Stipa breviflora; the most dominant species in arid and semiarid grassland) using open-top chambers (OTCs). The relationship between the N_mass of these four Stipa species and precipitation well fits a logarithmic function. The sensitivity of these four species to precipitation change was ranked as follows: S. bungeana > S. breviflora > S. baicalensis > S. grandis. The N_mass of S. bungeana was the most sensitive to precipitation change, while S. grandis was the least sensitive among these Stipa species. Elevated CO₂ exacerbated the effect of precipitation on N_mass. N_mass decreased under elevated CO₂ due to growth dilution and a direct negative effect on N assimilation. Elevated CO₂ reduced N_mass only in a certain precipitation range for S. baicalensis (163–343 mm), S. bungeana (164–355 mm), S. grandis (148–286 mm), and S. breviflora (130–316 mm); severe drought or excessive rainfall would be expected to result in a reduced impact of elevated CO₂. Elevated CO₂ affected the N_mass of S. grandis only in a narrow precipitation range. The effect of elevated CO₂ reached a maximum when the amount of precipitation was 253, 260, 217, and 222 mm for S. baicalensis, S. bungeana, S. grandis, and S. breviflora, respectively. The N_mass of S. grandis was the least sensitive to elevated CO₂. The N_mass of S. breviflora was more sensitive to elevated CO₂ under a drought condition compared with the other Stipa species.     

Phylogenetic Study of Methanol Oxidizers from Chilika-Lake Sediments Using Genomic and Metagenomic Approaches

Group-wise diversity of sediment methylotrophs of Chilika lake (Lat. 19°28′–19°54′N; Long. 85°06′–85°35′E) Odisha, India at various identified sites was studied. Both the culturable and unculturable (metagenome) methylotrophs were investigated in the lake sediments employing both mxaF and 16S rRNA genes as markers. ARDRA profiling, 16S rRNA gene sequencing, PAGE profiling of HaeIII, EcoRI restricted mxaF gene and the mxaF gene sequences using culture-dependent approach revealed the relatedness of α-proteobacteria and Methylobacterium, Hyphomicrobium and Ancyclobacter sp. The total viable counts of the culturable aerobic methylotrophs were relatively higher in sediments near the sea mouth (S3; Panaspada), also demonstrated relatively high salinity (0.1 M NaCl) tolerance. Metagenomic DNA from the sediments, amplified using GC clamp mxaF primers and resolved through DGGE, revealed the diversity within the unculturable methylotrophic bacterium Methylobacterium organophilum, Ancyclobacter aquaticus, Burkholderiales and Hyphomicrobium sp. Culture-independent analyses revealed that up to 90 % of the methylotrophs were unculturable. The study enhances the general understandings of the metagenomic methylotrophs from such a special ecological niche.

Electronic supplementary material

The online version of this article (doi:10.1007/s12088-015-0510-3) contains supplementary material, which is available to authorized users.     

Enhanced shoot multiplication in Ficus religiosa L. in the presence of adenine sulphate,glutamine and phloroglucinol

Ficus religiosa (Pipal) is a long-lived valuable multipurpose forest tree. The tree is exploited because of its religious, ornamental and medicinal value and the regeneration rate in natural habitat is low. An in vitro propagation protocol has been developed from nodal segments obtained from a 45–50-year old tree. The highest bud break frequency (100 %) followed by maximum number of multiple shoots (13.9) as well as length (2.47 cm) were obtained on Woody Plant medium (WPM) supplemented with 1.0 mg/l BAP along with 0.5 mg/l IAA. Two modifications in this medium resulted in enhanced shoot regeneration-one with 200 mg/l glutamine + 150 mg/l ADS (called as MM-1) giving 32.5 shoots per nodal explant while another modification—with 200 mg/l glutamine + 150 mg/l ADS + 100 mg/l phloroglucinol (called as MM-2) giving 35.65 shoots per explant. These two media were used for sub-culturing of shoots for 4 months. The rate of shoot multiplication was same during the first three sub-cultures on MM-1 and the shoots regenerated were healthy, afterwards shoot multiplication declined. While on MM-2, shoot multiplication declined after first sub-culture and shoots underwent the problem of early leaf fall. Rooting was best induced in micro-shoots excised from proliferated shoot cultures on semi-solid as well as liquid WPM modified with 2.0 mg/l IBA and 0.5 mg/l IAA. The in vitro-raised plantlets were potted and acclimatized under culture room conditions for 25–30 days before transfer to soil conditions, where the established plants showed more than 90 % survival.