Allelopathic effect of actinobacterial isolates against selected weeds

The taxonomic characteristics of weeds such as morphology of shoot, leaves, flowers, stem, fruits and seeds were recorded as Gynandropsis pentaphylla, DC, Amarantus spinosus Linn, Cyperus rotundus, Amarantus viridis Linn, Cassia occidentalis, Linn and Echinochilora orygicola. Totally 20 actinobacteria isolates were screened for herbicidal activity against the weed. Among the 20 isolates, only four actinobacterial isolates KA1-3, KA1-4, KA1-7 and KA23A showed significant herbicidal activity against C. rotundus. The herbicidal effect of actinobacterial culture filtrates on germination and seedling growth of C. rotundus was tested. The shoot and root growth of C. rotundus was severely affected when compared to control. The potent actinobacterial isolates KA1-4 and KA1-7 were characterised based on their morphological and molecular phylogenetic property and were identified as Streptomyces sp. The present study concludes that actinobacterial isolates will be used as bioherbicide against C. rotundus. Further studies are required to confirm the activity of actinobacterial isolates against C. rotundus under field conditions.     

Design,Synthesis, and Structure-Activity Relationship (SAR) Studies of Ketone-Isobenzofuranone Hybrid Herbicides

Thirty-five ketone-isobenzofuranone hybrids ( 1 – 35 ) were designed, synthesized, and evaluated for their herbicidal activity against Chinese amaranth (Amaranthus tricolor) and barnyard grass (Echinochloa crus-galli). The structure-activity relationship (SAR) results revealed that the position and type of substituent were crucial for activity. The o-substituted derivatives outperformed the m- and p-substituted derivatives. Compounds with strong electron-donating groups (OH, OMe) had low activity, while those with heterocycles (N-methylpyrrole, furan, and thiophene) had a moderate herbicidal effect. Compounds with a weak electron-donating group (Me) and weak, moderate, and strong electron-withdrawing groups (F, Cl, Br, and NO₂) showed promising herbicidal activity. Among these, the o-F substituted compound ( 20 ) was the most effective against Chinese amaranth, and the o-Cl substituted compound ( 23 ) was the most potent against barnyard grass. This is the first time the herbicidal potential of ketone-isobenzofuranone hybrids has been studied. The discovery of current chemical clues would be beneficial for the development of novel herbicides.     

CCC and Prohexadione-Ca Enhance Rhizome Growth and Lateral Bud Production in Rhubarb (<Emphasis Type="Italic">Rheum rhabarbarum</Emphasis> L.)

Accelerating rhizome growth is crucial to enhancing propagule production in rhubarb (Rheum rhabarbarum L.) because the crop is propagated through rhizome divisions. This can be achieved through manipulating source-sink activity. This study tested the hypothesis that synthetic plant growth retardants Prohexadione-Ca and CCC enhance rhizome growth in rhubarb. Two different concentrations of these plant growth retardants and GA₃ (positive control) were foliarly applied on the cultivar German Wine at three stages of shoot growth under greenhouse conditions. Both Prohexadione-Ca and CCC favorably enhanced rhizome growth through suppressing shoot growth. CCC at 3000 mg L⁻¹ produced the best results and the effect was apparent when applied at 12 weeks after shoot emergence. The rhizome diameter, fresh weight, and the number of viable buds were enhanced significantly in plants sprayed with CCC 3000 mg L⁻¹. Both Prohexadione-Ca and CCC were equally effective in enhancing dry mass and starch allocation preferentially toward the rhizome. Prohexadione-Ca- and CCC-induced rhizome growth enhancement could possibly be due to their known role as GA biosynthesis inhibitors or through increasing photosynthetic efficiency and preferentially reallocating carbohydrates to the rhizome.     

5种杀虫剂对西花蓟马和花蓟马的毒力及其生理酶活性的影响

【目的】探究不同杀虫剂对重要入侵害虫西花蓟马及其本地近缘种花蓟马的毒力及对保护酶和解毒酶活性的影响,为进一步研究2种害虫的抗性管理提供依据。【方法】采用浸渍法测定5种田间常用杀虫剂对西花蓟马和花蓟马的毒力,并测定杀虫剂亚致死浓度(LC25)下2种蓟马体内保护酶和解毒酶活性的差异。【结果】不同杀虫剂对2种蓟马的毒力依次为:乙基多杀菌素甲维盐阿维菌素吡虫啉噻虫嗪,乙基多杀菌素对西花蓟马和花蓟马的LC_(50)分别为0.28和0.03 mg·L~(-1)。不同药剂的亚致死剂量(LC_(25))对西花蓟马和花蓟马体内保护酶和解毒酶活性普遍具有诱导作用。其中,阿维菌素对西花蓟马超氧化物歧化酶(SOD)活性诱导作用最强,为326.40 U·mg~(-1),是对照的9.37倍,而乙基多杀菌素对花蓟马SOD活性诱导作用最强,为245.35 U·mg~(-1),是对照的9.32倍;吡虫啉对西花蓟马和花蓟马过氧化物酶(POD)诱导作用最强,分别为298.67和246.79 U·mg~(-1),是对照的37.10和20.57倍;阿维菌素对西花蓟马和花蓟马过氧化氢酶(CAT)和羧酸酯酶(CarE)诱导作用最强,分别为298.67、246.79 U·mg~(-1)(CAT活性)和12.53、11.99 U·mg~(-1)(CarE活性);乙基多杀菌素对西花蓟马和花蓟马谷胱甘肽转移酶(GST)和乙酰胆碱酯酶(AChE)诱导作用最强,分别为77527.59、66927.39 U·mg~(-1)(GST活性)和2.34、2.22 U·mg~(-1)(AChE活性)。【结论】5种杀虫剂中,乙基多杀菌素对2种蓟马的毒力最强;西花蓟马对杀虫剂的解毒代谢能力强于花蓟马。     

Phytochemical, antimicrobial, antioxidant and antigenotoxic potentials of Cyperus rotundus extracts

The aqueous, ethyl acetate, methanolic and Total Oligomer Flavonoids (TOF) enriched extracts, obtained from the aerial parts of Cyperus rotundus, were investigated for their contents in phenolic compounds. Antioxidative activity using the NBT/riboflavin assay system, antimicrobial activity against Gram positive and Gram negative bacterial reference strains as well as antigenotoxic activity tested with the SOS chromotest assay were also studied. Significant antibacterial activity against reference strains; Staphylococcus aureus, Enterococcus faecalis, Salmonella enteritidis and Salmonella typhimurium, was detected in the presence of ethyl acetate and TOF enriched extracts. In addition to their antimicrobial activity, the same extracts showed a significant ability to inhibit nitroblue tetrazolium reduction by the superoxide radical in a non enzymatic O₂^.− generating system, and were also able to reduce significantly the genotoxicity induced by nifuroxazide and Aflatoxin B1. The antioxidant, antimicrobial and antigenotoxic activities exhibited by C. rotundus depend on the chemical composition of the tested extracts.     

Contribution of plants to N 2 O emissions in soil-winter wheat ecosystem: pot and field experiments

Outdoor pot and field experiments were conducted to assess the role of growing plants in agricultural ecosystem N₂O emissions. N₂O emissions from plants were quantified as the difference in soil-crop system N₂O emissions before and immediately after cutting plants during the main growth stages in 2001–02 and 2002–03 winter wheat seasons. Emissions of N₂O from plants depended on biomass within the same plant developmental status. Field results indicated that the seasonal contribution of N₂O emissions from plants to ecosystem fluxes averaged 25%, ranging from 10% at wheat tillering to 62% at the heading stage. The fluxes of N₂O emissions from plants varied between 0.3 and 3.9 mg N₂O-N m⁻² day⁻¹ and its seasonal amount was equivalent to 0.23% of plant N released as N₂O. A N₂O emission coefficient (N₂O_E, mg N₂O-N g⁻¹ C day⁻¹), defined as N₂O-N emission in milligrams from per gram carbon of plant dry matter within a day, was represented by a 5-fold variation ranging from 0.021 to 0.004 mg N₂O-N g C⁻¹ day⁻¹. A linear relationship (y=0.4611x+0.0015, r ²=0.9352, p < 0.001) between N₂O_E (y) and plant dark respiration rate (x, mg CO₂-C g C⁻¹ day⁻¹) suggested that in the absence of photosynthesis, some N₂O production in plant N assimilation was associated with plant respiration. Although this study could not show whether N₂O was produced or transferred by winter wheat plants, these results indicated an important role for higher plant in N₂O exchange. Identifying its potential contribution is critical for understanding agricultural ecosystem N₂O sources.     

A New Apparatus for Preparative Gel Electrophoresis

Syntheses of two carbonyl derivatives of hydantocidin 1, a potent, naturally occurring herbicide, and their herbicidal activity are described. Spiroimidazolidinone 2, the descarbonyl compound at C9, was prepared by employing reductive demethylsulfurization with tri-n-butyltin hydride as the key step. Another derivative, spiroimidazolinone 10, was obtained from α-azidoamide 8 and benzyl isocyanate via the aza-Wittig reaction. 2 had lost almost all herbicidal activity, whereas 10 retained herbicidal activity against such dicotyledonous weeds as ragweed and cocklebur, but lost activity against monocotyledonous weeds. These results imply the possibility that proper modification of the carbonyl group at C7 of the parent compound would afford hydantocidin analogues possessing crop selectivity.     

施用生物炭和秸秆还田对华北农田CO2、N2O排放的影响

以华北农田冬小麦-夏玉米轮作体系连续6a施用生物炭和秸秆还田的土壤为研究对象,于2013年10月—2014年9月,采用静态暗箱-气相色谱法,对CO_2、N_2O通量进行了整个轮作周期的连续观测,探究施用生物炭与秸秆还田对其排放通量的影响。试验共设4个处理:CK(对照)、C1(低量生物炭4.5 t hm~(-2)a~(-1))、C2(高量生物炭9.0 t hm~(-2)a~(-1))和SR(秸秆还田straw return)。结果表明:在整个轮作周期内,各处理CO_2、N_2O通量随时间的变化趋势基本一致。随着生物炭施用量的增加,CO_2排放通量分别增加了0.3%—90.3%(C1)、1.0%—334.2%(C2)和0.4%—156.3%(SR)。其中,C2处理对CO_2累积排放量影响最大,增幅为42.9%。对N_2O而言,C2处理显著降低了N_2O累积排放量,但增加了CO_2和N_2O排放的综合增温潜势,C1和SR处理对N_2O累积排放量及综合增温潜势均没有显著影响。相关分析表明,土壤温度和土壤含水量是影响CO_2通量最主要的因素,两者之间呈极显著的正相关关系;N_2O通量与土壤温度、土壤含水量、NO_3~--N和NH_4~+-N均表现出极显著的正相关关系,而与土壤p H值表现出极显著的负相关关系。由此可见,添加生物炭对于减少氮素的气体损失具有较大的潜力。     

The synthesis and biological properties of 8-azido-N 6-benzyladenine,a potential photoaffinity reagent for cytokinin

A cytokinin photoaffinity reagent, 8-azido-N ⁶-benzyladenine (8N₃BA), was synthesized from 8-bromoadenosine via azide replacement, benzylation at N–1, rearrangement to the N-6-benzyl derivative and acid hydrolysis. The compound thus obtained was found to have full cytokinin activity in the moss and tobacco cell-suspension bioassays. Photolysis of 8N₃BA was accomplished with long and short-wavelength ultraviolet light and produced compounds which had very little or no biological activity in the two bioassays. In-vivo photolysis of 8N₃BA caused loss of the cytokinin activity of this compound in moss protonemata. This result was similar to earlier ones where the biological response of moss protonemata to benzyladenine was reversed following removal of the hormone by a short rinse with water.Abbreviations BA N ⁶-benzyladenine - 8N₃BA 8-azido-N ⁶-benzyladenine - PMR proton magnetic resonance - TLC thin-layer chromatography - UV ultraviolet In partial fulfillment of requirements for the Ph.D. degree at Michigan State University     

Nitrogenase activity in the papyrus swamps of Lake Naivasha,Kenya

Nitrogenase activity (acetylene reduction activity) was found to occur universally in the Cyperus papyrus swamp in Lake Naivasha. Low rates of acetylene reduction activity (0.9–104.9 nmol C₂H₄ g d.wt. roots^-1 h^-1) were associated with excised roots of C. papyrus but higher rates of activity (89.0–280.4 nmol C₂H₄ g d.wt. roots^-1 h^-1) were associated with intact root systems of the plant. It was estimated that nitrogen fixation associated with young roots alone could supply about 26% of the nitrogen requirements of growing papyrus plants. Acetylene reduction activity in the lake bottom sediments was generally low and associated with adjacent papyrus stands. Plate counts of putative aerobic and facultatively anaerobic N₂-fixing bacteria associated with papyrus roots showed the presence of high numbers of diazotrophs (5.4 × 10⁶ CFU g d.wt. roots^-1). Fewer numbers of N₂-fixing bacteria were detected in the sediments (1.9 × 10³-3.2 × 10⁴ CFU g d.wt. sediment^-1).     

N1, N14-diferuloylspermine as an antioxidative phytochemical contained in leaves of Cardamine fauriei

Most Brassicaceae vegetables are ideal dietary sources of antioxidants beneficial for human health. Cardamine fauriei (Ezo-wasabi in Japanese) is a wild, edible Brassicaceae herb native to Hokkaido, Japan. To clarify the main antioxidative phytochemical, an 80% methanol extraction from the leaves was fractionated with Diaion® HP-20, Sephadex® LH-20, and Sep-Pak® C18 cartridges, and the fraction with strong antioxidant activity depending on DPPH method was purified by HPLC. Based on the analyses using HRESIMS and MS/MS, the compound might be N¹, N¹⁴-diferuloylspermine. This rare phenol compound was chemically synthesized, whose data on HPLC, MS and ¹H NMR were compared with those of naturally derived compound from C. fauriei. All results indicated they were the same compound. The radical-scavenging properties of diferuloylspermine were evaluated by ORAC and ESR spin trapping methods, with the diferuloylspermine showing high scavenging activities of the ROO^·, O₂^·?, and HO^· radicals as was those of conventional antioxidants.     

Evaluation of Pomelo Seed Extracts as Natural Antioxidant,Antibacterial, Herbicidal Agents,and Their Functional Components

Pomelo seeds (PS) are important by-product of pomelo fruits (Citrus grandis Osbeck). The value-added utilization of PS remains highly challenged. This study aimed to investigate the utilization potential of PS as natural antioxidant, antibacterial, herbicidal agents, and their functional components. The ethanolic extract (EE) of PS and its four fractions as PEE (petroleum ether extract), AcOEtE (ethyl acetate extract), BTE (butanol extract), and WE (water extract), were prepared and biologically evaluated. BTE exhibited the best antioxidant activity among all these extracts, in both ABTS (2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) and FRAP (ferric reducing antioxidant power) assays. AcOEtE was superior to other extracts in herbicidal assay against both Festuca elata Keng (IC₅₀ of 0.48 mg mL⁻¹) and Amaranthus retroflexus L. (IC₅₀ of 0.94 mg mL⁻¹). Meanwhile, both AcOEtE and BTE demonstrated inhibitory effects against Bacillus subtilis, Escherichia coli, and Xanthomonas citri subsp. citri, with MIC ranging 2.5–5.0 mg mL⁻¹. Furthermore, the primary chemical components involving naringin, deacetylnomilin, limonin, nomilin, and obacunone, were quantified in all these extracts. PCA (principal component analysis) suggested that naringin might highly contribute to the antioxidant activity of PS, and the herbicidal activity should be ascribed to limonoids. This study successfully identified AcOEtE and BTE as naturally occurring antioxidant, antibacterial, and herbicidal agents, showing application potential in food and cosmetics industries, and organic farming agriculture.     

Two N 5, N 10-methylenetetrahydromethanopterin dehydrogenases in the extreme thermophile Methanopyrus kandleri: characterization of the coenzyme F420-dependent enzyme

It was recently reported that the extreme thermophile Methanopyrus kandleri contains only a H₂-forming N ⁵, N ¹⁰-methylenetetrahydromethanopterin dehydrogenase which uses protons as electron acceptor. We describe here the presence in this Archaeon of a second N ⁵,N ¹⁰-methylenetetrahydromethanopterin dehydrogenase which is coenzyme F₄₂₀-dependent. This enzyme was purified and characterized. The enzyme was colourless, had an apparent molecular mass of 300 kDa, an isoelectric point of 3.7±0.2 and was composed of only one type of subunit of apparent molecular mass of 36 kDa. The enzyme activity increased to an optimum with increasing salt concentrations. Optimal salt concentrations were e.g. 2 M (NH₄)₂SO₄, 2 M Na₂HPO₄, 1.5 M K₂HPO₄, and 2 M NaCl. In the absence of salts the enzyme exhibited almost no activity. The salts affected mainly the V _max rather than the K _m of the enzyme. The catalytic mechanism of the dehydrogenase was determined to be of the ternary complex type, in agreement with the finding that the enzyme lacked a chromophoric prosthetic group. In the presence of M (NH₄)₂SO₄ the V _max was 4000 U/mg (k _cat=2400 s^-1) and the K _m for N ⁵,N ¹⁰-methylenetetrahydromethanopterin and for coenzyme F₄₂₀ were 80 M and 20 M, respectively. The enzyme was relatively heat-stable and lost no activity when incubated anaerobically in 50 mM K₂HPO₄ at 90°C for one hour. The N-terminal amino acid sequence was found to be similar to that of the F₄₂₀-dependent N ⁵, N ¹⁰-methylenetetrahydromethanopterin dehydrogenase from Methanobacterium thermoautotrophicum, Methanosarcina barkeri, and Archaeoglobus fulgidus.Abbreviations H₄MPT tetrahydromethanopterin - F₄₂₀ coenzyme F₄₂₀ - CH₂=H₄MPT N ⁵,N ¹⁰-methylenetrahydromethanopterin - CHH₄MPT⁺ N ⁵,N ¹⁰-methenyltetrahydromethanopterin - methylene-H₄MPT dehydrogenase N ⁵,N ¹⁰-methylenetetrahydromethanopterin dehydrogenase - Mops N-morpholinopropane sulfonic acid - Tricine N-[Tris(hydroxymethyl)-methyl]glycine - 1 U = 1 mol/min     

Substituent Effect on Photosynthesis of N2-α-Substituted Benzyl-N4-alkyl-2,4-diamino-6-chloro-s-triazines and Their Phytotoxic Property

The influence of substituents on the activities of a series of N²-α-substituted benzyl-N⁴-alkyl-2,4-diamino-6-chloro-s-triazines as inhibitors of photosystem II (PSII) was examined, and the phytotoxic differences between them and atrazine, as to the photosynthesis in leaf disks, mesophyll cells, intact chloroplasts and broken chloroplasts of spinach, and as to seedling-growth, were discussed. The inhibitory activity of the N²-α,α-dimethylbenzyl-N⁴-ethyl derivative (6), which was comparable on that of atrazine, was lower than those of the N²-α-alkylbenzyl analogues (1 ~5). The N⁴-?-alkyl-N²-α- methylbenzyl derivatives, in spite of the carbon length of the alkyl group, exhibited more potent activity than atrazine, but an a α β substitution of the N⁴-n-alkyl group caused a decrease in the activity with a few exceptions. These data may imply that the space of the binding site on PSII surrounding both the N² and N⁴ amino groups is relatively large. The binding between the receptor site and the N⁴ amino group, however, is easily influenced by a slight structural change in an inhibitor. The herbicidal compounds, N²-α-methylbenzyl-A^⁴-ethyl (1), A^²-α,α-dimethylbenzyl-N⁴-1-methylpropyl (30) and N²-α-methylbenzyl-N⁴,N⁴-diethyl (42) derivatives, exhibited potent inhibitory activity in the seedling growth test under dark/light conditions, whereas atrazine was very poor. The inhibitory activity of compound (1) toward photosynthesis was poor with leaf disks, compared to atrazine, whereas, the order of their activities was the reverse for plant preparations such as abaxial epidermis peeled leaf disks, mesophyll cells, intact chloroplasts and broken chloroplasts. It was indicated that a change in the phytotoxic symptom in the whole plant assay would be correlated to the permeability of the compound through the plant membrane(s).     

Photosynthetic responses of apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) to photosynthetic photon flux density,leaf temperature,and CO<Subscript>2</Subscript> concentration

Two cultivars (Katy and Erhuacao) of apricot (Prunus armeniaca L.) were evaluated under open-field and solar-heated greenhouse conditions in northwest China, to determine the effect of photosynthetic photon flux density (PPFD), leaf temperature, and CO₂ concentration on the net photosynthetic rate (P _N). In greenhouse, Katy registered 28.3 μmol m⁻² s⁻¹ for compensation irradiance and 823 μmol m⁻² s⁻¹ for saturation irradiance, which were 73 and 117 % of those required by Erhuacao, respectively. The optimum temperatures for cvs. Katy and Erhuacao were 25 and 35 °C in open-field and 22 and 30 °C in greenhouse, respectively. At optimal temperatures, P _N of the field-grown Katy was 16.5 μmol m⁻² s⁻¹, 21 % less than for a greenhouse-grown apricot. Both cultivars responded positively to CO₂ concentrations below the CO₂ saturation concentration, whereas Katy exhibited greater P _N (18 %) and higher carboxylation efficiency (91 %) than Erhuacao at optimal CO₂ concentration. Both cultivars exhibited greater photosynthesis in solar-heated greenhouses than in open-field, but Katy performed better than Erhuacao under greenhouse conditions.     

A 30% reduction in switchgrass rhizome reserves did not decrease biomass yield

A long-standing question in perennial grass breeding and physiology is whether yield improvement strategies could compromise winter survival. Since perennial grasses rely on stored carbohydrates for winter maintenance and spring regrowth, yield improvement strategies could reduce winter survival if they increase biomass and grain yields at the expense of carbon allocation to storage. Therefore, it is crucial to comprehend the dependence of regrowth on storage reserves. We experimentally depleted switchgrass (Panicum virgatum L.) rhizome reserves by storing rhizomes for 2 weeks at 5°C (control treatment) and 25°C (reserve-depleted treatment). During the storage period rhizome respiration was 5.3× higher at 25°C (0.010 μmol CO₂ g⁻¹ min⁻¹ at 5°C vs. 0.054 μmol CO₂ g⁻¹ min⁻¹ at 25°C; p < 0.0001) and the starch content was depleted by 30% by the end of storage. Surprisingly, reserve-depleted switchgrass had 60% larger leaf area (LA; LA_control = 149 cm² pot⁻¹ vs. LA_depleted = 239 cm² pot⁻¹; p = 0.013) and produced ~40% more aboveground biomass than control plants (9.46 g pot⁻¹ vs. 6.63 g pot⁻¹; p = 0.112). In addition, reserve-depleted switchgrass restored its rhizome starch reserves to pre-storage levels. Switchgrass showed a large plasticity among its source-sink components to buffer the imposed reserve depletion. It increased plant photosynthesis by increasing the photosynthetic leaf area while keeping photosynthesis constant on a leaf area basis and readjusted the timing and activity of sink organs. These results suggest that switchgrass, and potentially other perennial grasses, largely over-invest in storage reserves. Therefore, current breeding strategies in perennial grasses aimed to extend the aboveground growing season should not compromise crop persistence. Our study also has implications on long-term yield dynamics as it highlights sink limitations as potential driver of the yield decline commonly observed in perennial grasses 5+ years after cultivation.     

Nitrogen fixation in coniferous bark litter

Summary Non-symbiotic heterotrophic N₂ fixation in coniferous bark litter was investigated with the acetylene reduction assay under aerobic and anaerobic conditions. The litter studied was composed essentially of bark, of pH 5 and a C/N ratio of 101; the ratio of available C to available N, which governs N₂ fixation, was considerably higher. The rate of N₂ fixation was estimated as 2.5–4.4 g N. g^–1 dry wt. day^–1. Nitrogenase activity was still evident after seven months of incubation under aerobic conditions. The N₂-ase activity was O₂ dependent: under anaerobic conditions no N₂-ase activity was found unless a fermentable C source was added. The importance of N₂ fixation in N-poor litter for the maintenance of soil fertility is emphasized.     

Boron Deficiency Induced Changes in Translocation of 14CO2-Photosynthate Into Primary Metabolites in Relation to Essential Oil and Curcumin Accumulation in Turmeric (Curcuma longa L.)

Changes in leaf growth, net photosynthetic rate (P _N), incorporation pattern of photosynthetically fixed ¹⁴CO₂ in leaves 1–4 from top, roots, and rhizome, and in essential oil and curcumin contents were studied in turmeric plants grown in nutrient solution at boron (B) concentrations of 0 and 0.5 g m^-3. B deficiency resulted in decrease in leaf area, fresh and dry mass, chlorophyll (Chl) content, and P _N and total ¹⁴CO₂ incorporated at all leaf positions, the maximum effect being in young growing leaves. The incorporation of ¹⁴CO₂ declined with leaf position being maximal in the youngest leaf. B deficiency resulted in reduced accumulation of sugars, amino acids, and organic acids at all leaf positions. Translocation of the metabolites towards rhizome and roots decreased. In rhizome, the amount of amino acids increased but content of organic acids did not show any change, whereas in roots there was decrease in contents of these metabolites as a result of B deficiency. Photoassimilate partitioning to essential oil in leaf and to curcumin in rhizome decreased. Although the curcumin content of rhizome increased due to B deficiency, the overall rhizome yield and curcumin yield decreased. The influence of B deficiency on leaf area, fresh and dry masses, CO₂ exchange rate, oil content, and rhizome and curcumin yields can be ascribed to reduced photosynthate formation and translocation.     

Purification and properties of N 5 ,N 10 -methylenetetrahydromethanopterin reductase (coenzyme F420-dependent) from the extreme thermophile Methanopyrus kandleri

Methanopyrus kandleri belongs to a novel group of abyssal methanogenic archaebacteria that can grow at 110°C on H₂ and CO₂ and that shows no close phylogenetic relationship to any methanogens known so far. N ⁵ N ¹⁰ -Methylenetetrahydromethanopterin reductase, an enzyme involved in methanogenesis from CO₂, was purified from this hyperthermophile. The apparent molecular mass of the native enzyme was found to be 300 kDa. Sodium dodecylsulfate/polyacrylamide gel electrophoresis revealed the presence of only one polypeptide of apparent molecular mass 38 kDa. The ultraviolet/visible spectrum of the enzyme was almost identical to that of albumin indicating the absence of a chromophoric prosthetic group. The reductase was specific for reduced coenzyme F₄₂₀ as electron donor; NADH, NADPH or reduced dyes could not substitute for the 5-deazaflavin. The catalytic mechanism was found to be of the ternary complex type as deduced from initial velocity plots. V _max at 65°C and pH 6.8 was 435 U/mg (k_cat=275 s^-1) and the K _m for methylenetetrahydro-methanopterin and for reduced F₄₂₀ were 6 M and 4 M, respectively. From Arrhenius plots an activation energy of 34 kJ/mol was determined. The Q ₁₀ between 40°C and 90°C was 1.5.The reductase activity was found to be stimulated over 100-fold by sulfate and by phosphate. Maximal stimulation (100-fold) was observed at a sulfate concentration of 2.2 M and at a phosphate concentration of 2.5 M. Sodium-, potassium-, and ammonium salts of these anions were equally effective. Chloride, however, could not substitute for sulfate or phosphate in stimulating the enzyme activity.The thermostability of the reductase was found to be very low in the absence of salts. In their presence, however, the reductase was highly thermostable. Salt concentrations between 0.1 M and 1.5 M were required for maximal stability. Potassium salts proved more effective than ammonium salts, and the latter more effective than sodium salts in stabilizing the enzyme activity. The anion was of less importance.The N-terminal amino acid sequence of the reductase from M. kandleri was determined and compared with that of the enzyme from Methanobacterium thermoautotrophicum and Methanosarcina barkeri. Significant similarity was found.Abbreviations H₄MPT tetrahydromethanopterin - CH₂=H₄MPT N ⁵ ,N ¹⁰ -methylene-H₄MPT - CH₃-H₄MPT N ⁵-methyl-H₄MPT - CHH₄MPT⁺ N ⁵ ,N ¹⁰ -methenyl-H₄MPT - F₄₂₀ coenzyme F₄₂₀; 1 U=1 mol/min     

Dormancy break of celery (Apium graveolens L.) seeds by plant derived smoke extract

Seed dormancy of a highly-dormant cultivar of celery (Apium graveolens L.) was broken by combinations of plant-derived smoke extract or N⁶-benzyladenine (BA) and gibberellins A_4/7 (GA_4/7) in the dark at temperatures between 18 and 26°C. A less dormant cultivar which responded to GA_4/7 alone showed no additional response to smoke extract or BA. Neither smoke extract nor BA affected either cultivar in the dark in the absence of GA_4/7. The partial dormancy-breaking effect of short exposures to red-light was also enhanced by smoke extracts in this highly-dormant cultivar. The results suggest that smoke extracts act in a similar way to cytokinins, by enhancing gibberellin activity in the celery seed system.Abbreviations BA N⁶-benzyladenine - GA_4/7 A₄ and A₇ gibberellin mixture