水杨酸对真菌诱导子诱导红豆杉悬浮细胞膜脂过氧化和紫杉醇生物合成的影响

研究了 5 0 mg· L- 1真菌诱导子 (F5) ,5 0 mg· L- 1水杨酸 (SA) ,5 0 mg· L- 1F5+5 0 mg· L- 1SA3种处理 ,对红豆杉悬浮细胞膜脂过氧化和紫杉醇合成的影响。结果表明 :F5和 SA单独处理红豆杉细胞均引起细胞膜脂过氧化。SA+F5联合处理可以减轻 F5单独处理细胞所引起的膜脂过氧化程度 ,SA+F5联合处理与真菌诱导子处理相比 ,较大地提高了过氧化物酶的活性 ,得到较多的生物量。3种处理方法均可提高红豆杉细胞紫杉醇产量 ,特别以 F5+SA处理得到产量最高 ,达到 1 1 .5 mg· L- 1,分别为 F5,SA和对照组的 1 .5倍、2 .0倍和7.5倍。结果显示 :在真菌诱导子诱导与水杨酸的联合作用下提高紫杉醇产量 ,可能与水杨酸减轻真菌诱导子所引起的细胞膜脂过氧化程度有关     

Differential responses of soybean genotypes to excess manganese in an acid soil

To determine the tolerance of soybean genotypes to Mn toxicity, a green house study was conducted. Hayesville sandy loam (clayey, oxidic, mesic, Typic Hapludult), high in manganese, was used for the experiment. The experimental design was split-plot with three replications. Forty-one different soybean genotypes were planted in pots at two different pH levels: 5.2 (original soil pH) and 6.4 (amended with lime). Soybean genotypes were allowed to grow to the dry pod stage.Soil pH levels affected the soybean genotypes yields significantly (p < 0.01). Tolerant genotypes showed a higher or similar seed yield at pH 5.2 compared to pH 6.4. Sensitive genotype yields were lower at pH 5.2 than at pH 6.4. In general, Mn in leaves was higher at pH 5.2 than at pH 6.4. Some of the sensitive genotypes at pH 5.2. showed severe chlorosis and crinkle leaf symptoms as a result of Mn toxicity. Excess available Mn at pH 5.2. induced Ca deficiency. Soybean genotypes PI423758, PI417440, Aoda, Kingston, Rokusum and some others were tolerant to Mn toxicity, whereas PI417288, Verde, Wilson 5, Sango, Funk Delicious and some others were sensitive to Mn toxicity. The genotypes found to be tolerant can be recommended to plant breeders for development of Mn-tolerant cultivars.     

Induction by light of hydroxycinnamoyl-CoA: Quinate hydroxycinnamoyl transferase in buckwheat (Fagopyrum esculentum Moench): Absence of feed-forward control by trans-cinnamate

The light-induced increase in the activity of hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase (CQT) in hypocotyls of etiolated buckwheat seedlings is not suppressed by aminooxy acetic acid, an inhibitor of cinnamic acid formation in this tissue. Incubation of hypocotyls in 2 mmol l^-1 trans-cinnamate in the dark does not increase CQT activity. Thus, trans-cinnamate does not appear to mediate the effect of light on CQT in buckwheat hypocotyls.Abbreviations AOA aminooxy acetic acid - CA4H cinnamic acid-4-hydroxylase - CQT hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase - PAL L-phenylalanine ammonialyase     

Alginic acids in Lessonia vadosa: Partial hydrolysis and elicitor properties of the polymannuronic acid fraction

Extraction of Lessonia vadosa(Laminariales, Phaeophyta) collectedin three different localities near PuntaArenas in the south of Chile, with 3%aqueous sodium carbonate solutions gave asodium alginate yield in the range3.0–17.7% dry weight. There were markeddifferences in the mannuronic acid toguluronic acid ratio (M/G) (0.21–1.69) inthe alginic acids, samples collected inPuerto del Hambre in winter were composedmainly of mannuronic acid residues. Thealginate samples were characterized bypartial hydrolysis and FT-IR spectroscopy.No relationship was found between tissuetype and polyguluronic acid content. Theelicitor activity of the polymannuronicacid enriched fraction from alginic acid ofblades from Puerto del Hambre was assayedin wheat plants. The polymannuronic acidenriched fraction induced substancialelicitation of phenylalanine ammonia-lyase(PAL) and peroxidase (POD) activities.     

Effect of yeast elicitor and salicylic acid on the fluctuation of phytohormone contents in Ti-transformed Salvia miltiorrhiza cell cultures

When Ti transformed Salvia miltiorrhiza cells werecultured in a MS-NH₄ medium (MS without ammonium nitrate, containing30 g/L sucrose) at 25 °C in darkness for 18d, the total tanshinone (cryptotanshinone and tashinone IIA)contents in cultures were 12.23 mg/L and 15.07 mg/Lfor yeast elicitor (4 g/L), and yeast elicitor plus 200mol/L salicylic acid (SA) treated cultures, respectively,whereas only trace amounts of tanshinone were detected in the control or SAtreated cells. To explore the hormonal background concerning these phenomena,endogenous phytohormones were determined using ELISA kits. We found that ABA andiPAs contents in yeast elicitor plus SA treated cell cultures were increased 2.8to 9.8-fold and 3.6 to 5.8-fold respectively, while contents of GA₁and IAA were decreased by 13.2%–56.9% and 34.8%–74.6% respectively.This suggests that higher levels of ABA and iPAs combined with lower levels ofGA₁ and IAA inhibit the growth of cells, then probably stimulate thetanshinone production.     

氮磷钾肥对有柄石韦生理及绿原酸合成积累的影响

为探讨氮磷钾3种养分对有柄石韦生理及有效成分绿原酸合成积累的影响,该研究以有柄石韦组培苗为材料,分别用低养分(不施肥:N₀,P₀,K₀)、正常施肥(N:0.20 g·kg^-1,P:0.15 g·kg^-1,K:0.15 g·kg^-1)和高养分(N₁:0.40 g·kg^-1,P₁:0.30 g·kg^-1,K₁:0.30 g·kg^-1)3个浓度梯度,设置7个处理分别为NPK、N₀PK、N₁PK、NP₀K、NP₁K、NPK₀、NPK₁,测定不同处理下有柄石韦的抗性生理指标、绿原酸(CGA)含量及其合成关键酶活性。结果表明:(1)氮磷钾肥对有柄石韦的抗性生理有显著的影响,超氧化物歧化酶(SOD)在高氮和低钾处理中活性显著增加,而3种养分的低浓度和高浓度处理均会导致过氧化氢酶(CAT)活性显著上升。(2)不同养分水平氮、磷和钾对有柄石韦CGA含量存在显著影响,正常施肥的CGA含量最高,达到12.92 mg·g^-1; 高钾施肥的CGA含量最低,为7.79 mg·g^-1; 钾肥对CGA含量影响最显著。(3)CGA合成关键酶活性在不同施肥处理中差异显著,CGA含量与奎宁酸羟基肉桂酰转移酶(HQT)和4-香豆酰辅酶A连接酶(4CL)活性呈显著正相关,与莽草酸羟基肉酰转移酶(HCT)活性显著负相关,HQT、4CL和HCT是导致CGA含量差异的关键因素。该研究结果为有柄石韦药材的人工栽培提供了理论依据。     

Interactive effects of methyl jasmonate and salicylic acid on floret opening in spikelets of Sorghum

Floret opening of excised spikelets in Sorghum bicolor (L.) Moench and Sorghum Sudanesis (Piper) Stapf was significantly stimulated by immersing into 2 mM methyl jasmonate (MeJA) solution. In male sterile (MS) lines of Sorghum bicolor (L.) Moench, floret opening was more sensitive to MeJA than that in maintainer (MT) lines. Salicylic acid (SA) could abolish the effect of MeJA on the opening of spikelets. These data indicate that MeJA plays a role in floret opening in sorghum, and that SA interacts with MeJA in regulating of this process.     

Studies on carbon flow in Crassulacean acid metabolism during the initial light period

In the Crassulacean acid metabolism (CAM) plants Kalanchoë tubiflora and Sedum morganianum a shift in the pathways occurs by which external CO₂ enters the metabolism during the initial light period (phase II of the diurnal CAM cycle). At the beginning of phase II, CO₂ is fixed mainly by the C₄ pathway; during late phase II, however, it is fixed mainly via the C₃ pathway. The C₃ pathway contributes to the phosphoenolpyruvate-carboxylase-mediated CO₂ fixation by the provision of three-carbon skeletons. Since the shift in the carbon-flow pathway is delayed after a CO₂-free night when malic-acid accumulation in the vacuoles is prevented, it is very likely that the amount of malic acid in the vacuole is integrated in the mechanism which controls CAM during the initial light period. A light-on signal at the beginning of phase II is not required to bring about the shifts in the carbon-flow pathways, as is shown by the reaction of plants to a prolonged dark period. A model of carbon flow during phase II is proposed.Abbreviations CAM Crassulacean acid metabolism - PEP-Case phosphoenolpyruvate carboxylase     

Influence of formic acid on fungal flora of barley and on aflatoxin production inAspergillus flavus link

In recent yearsAspergillus flavus and aflatoxin production have been noted on several occasions in grain preserved with formic acid. Samples of mouldy barley treated with formic acid and stored in an open bin were investigated for the presence of fungi. In the lower part of the bin there was a clear dominance ofFusarium sporotrichioides, and deoxynivalenol and neosolaniol were detected.A. flavus andA. fumigatus were also present.Paecilomyces variotii occurred, almost as a pure culture, in the upper part of the bin, but no patulin was found. Cultivation of four fungal isolates from these genera on laboratory substrates containing formic acid showedP. variotii to be the most tolerant to formic acid, withstanding 150 mM, but still without patulin production.F. sporotrichioides andA. fumigatus tolerated only 6 mM formic acid. The growth ofA. flavus was reduced and atypical at 60 mM formic acid. Pretreatment ofA. flavus spores with formic acid increased aflatoxin production about 800 times.     

Rapid changes in oxidative metabolism as a consequence of elicitor treatment of suspension-cultured cells of French bean (Phaseolus vulgaris L.)

Stressed plant cells often show increased oxygen uptake which can manifest itself in the transient production of active oxygen species, the oxidative burst. There is a lack of information on the redox status of cells during the early stages of biotic stress. In this paper we measure oxygen uptake and the levels of redox intermediates NAD/NADH and ATP and show the transient induction of the marker enzyme for redox stress, alcohol dehydrogenase. Rapid changes in the redox potential of elicitor-treated suspension cultures of French bean cells indicate that, paradoxically, during the period of maximum oxygen uptake the levels of ATP and the NADH/NAD ratio fall in a way that indicates the occurrence of stress in oxidative metabolism. This period coincides with the maximum production of active oxygen species particularly H₂O₂. The cells recover and start producing ATP immediately upon the cessation of H₂O₂ production. This indicates that the increased O₂ uptake is primarily incorporated into active O₂ species. A second consequence of these changes is probably a transient compromising of the respiratory status of the cells as indicated in expression of alcohol dehydrogenase. Elicitor-induced bean ADH was purified to homogeneity and the M_r 40 000 polypeptide was subjected to amino acid sequencing. 15% of the whole protein was sequenced from three peptides and was found to have nearly 100% sequence similarity to the amino acid sequence for pea ADH1 (PSADH1). The cDNA coding for the pea enzyme was used to demonstrate the transient induction of ADH mRNA in elicitor-treated bean cells. Enzyme activity levels also increased transiently subsequently. Increased oxygen uptake has previously been thought to be associated with provision of energy for the changes in biosynthesis that occur rapidly after perception of the stress signal. However the present work shows that this rapid increase in oxygen uptake as a consequence of elicitor action is not wholly associated with respiration.     

Rapid induction of ethylene biosynthesis in cultured parsley cells by fungal elicitor and its relationship to the induction of phenylalanine ammonia-lyase

The biosynthesis of ethylene was examined in suspension-cultured cells of parsley (Petroselinum hortense) treated with an elicitor from cell walls of Phytophthora megasperma. Untreated cells contained 50 nmol g^-1 of the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), and produced ethylene at a rate of about 0.5 nmol g^-1 h^-1. Within 2 h after addition of elicitor to the culture medium, the cells started to produce more ethylene and accumulated more ACC. Exogenously added ACC did not increase the rate of ethylene production in control or elicitor-treated cells, indicating that the enzyme converting ACC to ethylene was limiting in both cases. The first enzyme in ethylene biosynthesis, ACC synthase, was very rapidly and transiently induced by the elicitor treatment. Its activity increased more than tenfold within 60 min. Density labelling with ²H₂O showed that this increase was caused by the denovo synthesis of the enzyme protein. Cordycepin and actinomycin D did not affect the induction of ACC synthase, indicating that the synthesis of new mRNA was not required. The peak of ACC-synthase activity preceded the maximal phenylalanine ammonia-lyase (PAL) activity by several hours. Exogenously supplied ethylene or ACC did not induce PAL. However, aminoethoxyvinylglycine, an inhibitor of ACC synthase, suppressed the rise in ethylene production in elicitor-treated cells and partially inhibited the induction of PAL. Exogenously supplied ACC reversed this inhibition. It is concluded that induction of the ethylene biosynthetic pathway is a very early symptom of elicitor action. Although ethylene alone is not a sufficient signal for PAL induction, the enhanced activity of ACC synthase and the ethylene biosynthetic pathway may be important for the subsequent induction of PAL.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - PAL phenylalanine ammonia-lyase     

Effects of temperature and photosynthetic inhibitors on light activation of C4-phosphoenolpyruvate carboxylase

Phosphoenolpyruvate carboxylase from leaves of the C₄ plant Setaria verticillata (L.) Beauv. is activated by light; day levels of activity are reached after 30 minutes of illumination. Photoactivation is prevented by inhibitors of photosynthetic electron flow or of photophosphorylation and by D,L-glyceraldehyde, which inhibits the reductive pentose phosphate pathway.Although the extractable activity in the dark is not affected by temperature the photoactivation is prevented when both illumination and extraction are done under low temperature (5 C). High temperature (30 C) during either illumination or extraction is needed for activation. Once the enzyme is photoactivated at 30 C, a transfer of the leaves to 5 C does not abolish the extra activity.The results suggest that both unimpaired electron flow and photophosphorylation are prerequisites for the activation of phosphoenolpyruvate carboxylase. Low temperature apparently suppresses either the transport to the cytoplasm of a photosynthetic intermediate or the activating reaction itself. The inclusion of phosphoenolpyruvate in the extraction medium increases the night activity.On the basis of the available information, it is suggested that phosphoenolpyruvate could be the activator in vivo. In that case, the activation of phosphoenolpyruvate carboxylase would depend on internal CO₂ level and prior photoactivation of both pyruvate, orthophosphate, dikinase and NADP malate dehydrogenase.Abbreviations PEPCase phosphoenolpyruvate carboxylase - PEP phosphoenolpyruvate - PAR photosynthetically active radiation - CCCP carbonyl cyanide m-chlorophenylhydrazone - DCMU 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea - DSPD disalicylidenpropanediamine - MV methylviologen - ME malic enzyme - MDH malate dehydrogenase - PPDK pyruvate, Pi dikinase - CAM Crassulacean Acid Metabolism     

The effect of light quality and gibberellic acid (GA3) on photosynthesis and respiration rates of pea seedlings

CO₂ exchange were measured on pea seedlings (Pisum sativum L. var. Bördi) cultivated from seeds imbibed either in water (C-plants) or in gibberellic acid (GA₃) at the concentration of 25 g/1 (GA-plants), and then grown under 17 W/m² blue light (B-plants) or 11 W/m² red light (R-plants).When measured under the same light conditions as during growth the net photosynthesis (APS) rate in B-plants was about twice higher than that in R-plants. Dark respiration (DR) rate was 70% higher in B- than in R-plants. Red light retarded the development of photosynthetic activity, but GA₃ suppressed this effect. The hormone enhanced net photosynthesis and dark respiration to the same extent.When measured under saturating white light net photosynthesis rate of C-plants was also two times higher in B-plants than in R-plants. Growth conditions had only a slight effect on the APS of GA-plants under white light. APS rates of GA-plants grown under red light were higher under white light than those of C-plants, but lower than those of plants grown under blue light.We assume that blue light induced formation of plants that were adapted to higher light intensity: red light had an opposite effect, whereas gibberellic acid induced formation of plants that were adapted to medium light intensity.     

Differential induction of phenylalanine ammonia-lyase activity in date palm roots in response to inoculation with Fusarium oxysporum f. sp. albedinis and to elicitation with fungal wall elicitor

The inoculation of the roots of resistant (BSTN) and susceptible (JHL) cultivars of date palm seedlings byFusarium oxysporum f. sp.albedinis (Foa) induces an increase in activity of phenylalanine ammonia-lyase (E.C. 4. 3. 1. 5., PAL). The post-infectional response in the PAL activity in the resistant cultivar roots was faster and higher than that in the susceptible cultivar. However, the elicitation of the seedlings by the hyphal wall preparation (HWP) ofFoa induces an identical PAL response in the resistant and the susceptible cultivars. The elicitor activity of HWP was dose-dependent, the optimal concentration which induces a maximum PAL activity was 10 mg of mycelium per mL. The elicitor present in the HWP was thermostable since its elicitor activity was maintained after heat treatment (121 °C for 45 min). The treatment of the HWP with protease (Pronase E) does not have an effect on the HWP elicitor activity. However, the treatment of the HWP with sodium periodate inhibits its elicitor activity. This data suggests that the HWP elicitor is a carbohydrate compound. In addition, the HWP elicitor is non-specific since it induces identical responses of the PAL activity in two cultivars showing different behaviors to the pathogen. The absence of specificity of HWP elicitors and the differential response of the PAL activity to the infection byFoa and to the elicitation by the HWP are discussed. An explanation of the general interactions between plant and parasite is proposed.     

Differential effects of abscisic acid and methyl jasmonate on endoproteinases in senescing barley leaves

Endoproteinase activity was analyzed in chloroplasts isolated from barley leaf segments incubated in the dark with various hormonal senescence effectors. As a control, the endoproteinase activity of the supernatant fraction obtained during chloroplast preparation was also analyzed. Measured against azocaseine as substrate, the endoproteinase activity in chloroplasts increased 18 fold during the induction of senescence. This rise in activity was inhibited by kinetin (the activity increased only 10 fold) and very strongly stimulated by abscisic acid (ABA) (117 fold) and methyl jasmonate (Me-JA) (57 fold). Although less so, the endoproteinase activity of the supernatant fraction, mainly vacuolar and with acid pH optimum, was affected in the same way by all three effectors. Among the five endoproteinases (EC) found in chloroplasts, EC2 and EC4 were induced after incubation in water. ABA increased the levels of EC2 and EC4 (5 fold), and induced the development of EC3 and EC5, while Me-JA totally inhibited EC2 and EC4, and induced the development of EC1. At least one of the endoproteinases, EC2, is synthesized in chloroplasts. Among the six endoproteinases found in the supernatant fraction (E), E1, E2, E3 and E5, which are very probably extrachloroplastic endoproteinases, are stimulated by ABA to varying degrees. However, Me-JA stimulates E1 to a greater extent and totally inhibits E3. The differential effects of ABA and Me-JA on chloroplast and supernatant fraction endoproteinases suggest different action mechanisms for both senescence promotors.Abbreviations ABA abscisic acid - DTT dithiothreitol - E supernatant fraction endoproteinase - EC chloroplast endoproteinase - Me-JA methyl jasmonate - PNP p-nitrophenol - SDS-PAGE polyacrylamide gel electrophoresis containing sodium dodecyl sulphate - TCA trichloroacetic acid     

Photosynthetic acclimation of higher plants to growth in fluctuating light environments

This paper describes a study into the potential of plants to acclimate to light environments that fluctuate over time periods between 15 min and 3 h. Plants of Arabidopsis thaliana (L.) Heynh., Digitalis purpurea L. and Silene dioica (L.) Clairv. were grown at an irradiance 100 mol m^-2 s^-1. After 4–6 weeks, they were transferred to light regimes that fluctuated between 100 and either 475 or 810 mol m^-2 s^-1, in a regular cycle, for 7 days. Plants were shown, in most cases, to be able to undergo photosynthetic acclimation under such conditions, increasing maximum photosynthetic rate. The extent of acclimation varied between species. A more detailed study with S. dioica showed that this acclimation involved changes in both Rubisco protein and cytochrome f content, with only marginal changes in pigment content and composition. Acclimation to fluctuating light, at the protein level, did not fully reflect the acclimation to continuous high light - Rubisco protein increased more than would be expected from the mean irradiance, but less than expected from the high irradiance; cytochrome f increased when neither the mean nor the high irradiance would be expected to induce an increase.This revised version was published online in October 2005 with corrections to the Cover Date.     

Calcium and calmodulin inhibitor effects on the growth of carrot (Daucus carota L.) and radish (Raphanus sativus L.) in nutrient culture

Growth of carrot and radish seedlings in nutrient culture was inhibited by pretreatment with three calmodulin inhibitors. There was little selective effect on specific organs, shoots, tap root and fibrous roots over a range of concentrations. Although pretreatment with CaCl₂ (0.5 mM) did not affect growth of untreated seedlings, it partially reduced the inhibitory effects of trifluoperazine over the concentration range 0.01–0.05 mM. Trifluoperazine reduced the growth of GA₃-treated seedlings but did not overcome the modifying effect of GA₃ in favouring shoot/root ratio; ABA exacerbated its inhibitory effect on overall seedling growth and particularly on tap root development.Abbreviations GA₃ gibberellic acid - ABA abscisic acid - CaCl₂ calcium chloride - GAs gibberellins - Tfp trifluoperazine     

Inhibitory effects of chlorogenic acid and its related compounds on the invasion of hepatoma cells in culture

Actions of chlorogenic acid, a major component of coffee, andits constituents, caffeic and quinic acids, on theproliferation and invasion of AH109A, a rat ascites hepatomacell line, were investigated using in vitro assay systems. Allthree components suppressed the AH109A invasion atconcentrations of 5–40 M without altering the cellproliferation. At the concentration of 10 M, chlorogenic,caffeic and quinic acids significantly (P < 0.05) suppressedthe invasion by 68%, 36% and 31%, respectively, implying thatthe suppressive effect of chlorogenic acid on the AH109Ainvasion might result from the additive effects of itsconstituents, caffeic and quinic acids. At the concentrationof 10 M, cinnamic acid and p-coumaric acid (4-hydroxycinnamicacid) exerted no or little influence on the invasion, whereascaffeic acid (3,4-dihydroxycinnamic acid) significantly (P <0.05) suppressed it, suggesting the possible involvement ofthe 3,4-dihydroxy group of caffeic acid in the suppression.Chlorogenic acid was thus demonstrated to be one of thechemical entities in coffee suppressing the hepatoma invasionin vitro, and both of its constituents, caffeic and quinicacids, to be responsible for the anti-invasive activity. Theseresults suggest the existence of nutritionally andpharmacologically important substances in coffee which controltumor cell invasion.