Germination of Echinochloa crus-galli (Barnyard Grass) Seeds under Anaerobic Conditions : Respiration and Response to Metabolic Inhibitors

Echinochloa crus-galli L. Beauv., a rice-field weed, can germinate and grow for extended periods of time in an anaerobic environment. Compared to pea, which does not germinate under anaerobiosis, the evolution of CO₂ in Echinochloa and rice is lower and the peak rate of CO₂ evolution is delayed when germinated without oxygen. The plants studied also differ with respect to their respiration ratio ([CO₂] N₂/[CO₂] air) and metabolism used during the early stages of germination. Echinochloa does not increase its glycolytic rate under anaerobiosis, whereas pentose phosphate pathway activity appears to increase during the first 40 to 50 hours of germination.

Based on its response to metabolic inhibitors (NaF, dinitrophenol, and malonate), anaerobic metabolism in Echinochloa proceeds primarily through glycolysis, with partial operation of the tricarboxylic acid cycle and little or no oxidative phosphorylation. Also, Echinochloa is sensitive to CN during aerobic germination, whereas rice appears to be able to shift to CN-insensitive electron transport. Finally, the effectiveness of cyanide and azide on inhibiting germination of Echinochloa in N₂, but not CO, suggests that cytochrome oxidase is not used to reoxidize pyridine nucleotides in the absence of oxygen. The possible existence of an alternate electron acceptor is discussed.

     

代谢工程改造运动发酵单胞菌用于提高乙醇产量

目的:采用可以在运动发酵单胞菌中表达的操纵子构建重组运动发酵单胞菌,用于提高该细菌对高温高糖的耐受性和提高乙醇产量.方法:用外来的YfdZ、MetB和Hsp构建的多顺反子质粒,转化运动发酵单胞菌而使其获得新的代谢途径.在玉米水解液中,验证了该多顺反子质粒对运动发酵单胞菌产生乙醇的影响.结果:与对照菌相比,在37℃和糖浓度为28%的培养条件下,该基因工程菌的乙醇产量提高到183.2%.在37℃,糖浓度为28%并添加氮源的条件下,该基因工程菌的乙醇产量提高到148.0%.结论:YfdZ、MetB及Hsp三种基因的共同作用能显著提高运动发酵单胞菌的乙醇产量和发酵温度.     

Response of Soybean Seed Germination to Cadmium and Acid Rain

Cadmium (Cd) pollution and acid rain are the main environmental issues, and they often occur in the same agricultural region. Nevertheless, up to now, little information on the combined pollution of Cd(2+) and acid rain action on crops were presented. Here, we investigated the combined effect of Cd(2+) and acid rain on the seed germination of soybean. The results indicated that the single treatment with the low level of Cd(2+) (0.18, 1.0, 3.0?mg?L(-1)) or acid rain (pH ≥3.0) could not affect the seed germination of soybean, which was resulted in the increased activities of peroxidase and catalase. The single treatment with the high concentration of Cd(2+) (>6?mg?L(-1)) or acid rain at pH?2.5 decreased the activities of peroxidase and catalase, damaged the cell membrane and then decreased the seed germination of soybean. Meanwhile, the same toxic effect was observed in the combined treatment with Cd(2+) and acid rain, and the combined treatment had more toxic effect than the single treatment with Cd(2+) or acid rain. Thus, the combined pollution of Cd(2+) and acid rain had more potential threat to the seed germination of soybean than the single pollution of Cd(2+) or acid rain.     

GAPDH Expression Predicts the Response to R-CHOP,the Tumor Metabolic Status,and the Response of DLBCL Patients to Metabolic Inhibitors

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种子萌发期4种植物对干旱胁迫的响应及其抗旱性评价研究

用不同浓度PEG-6000的1/2Hoagland溶液模拟干旱胁迫条件,对沙枣、柠条、杠柳、白蜡4种植物种子的总萌发率、幼苗鲜重、活力指数进行研究.结果表明:(1)随着干旱胁迫程度的加剧4种植物种子的总萌发率、幼苗鲜重、活力指数呈下降趋势,且白蜡下降幅度最大,但柠条、沙枣、杠柳的抗旱指数显著高于白蜡;(2)4种植物种子的逐日萌发率和胚根日变化表明:低浓度PEG-6000对柠条和杠柳的种子萌发具有较好的诱发作用,白蜡和杠柳种子萌发比较集中,沙枣和柠条相对比较分散;沙枣、柠条和白蜡胚根生长呈线性增长,而杠柳呈指数式增长;(3)采用隶属函数法对干旱胁迫下4种植物的相对萌发率、相对鲜重、抗旱指数、相对活力指数进行综合评价的结果显示,4种植物种子萌发阶段的抗旱性次序为:柠条>沙枣>杠柳>白蜡.     

代谢工程改善野生酵母利用木糖产乙醇的性能

从256个自然样品中筛选得到1株可高效转化D-木糖的酵母。通过生理生化和分子生物学方法鉴定, 证实该菌株是属于Candida tropicalis。以该酵母为研究对象, 增加木糖醇脱氢酶表达量, 通过改变代谢流以达到提高酒精产率的目的。以pXY212-XYL2质粒为基础载体, 构建了含有潮霉素抗性的pYX212-XYL2-Hygro, 电击转化进入野生型C. tropicalis, 潮霉素抗性筛选, 得到含高拷贝木糖醇脱氢酶基因的重组菌株C. tropicalis XYL2-7。重组菌的比酶活达到0.5 u/mg protein, 比原始菌株提高了3倍。实验表明, 重组菌木糖醇得率比原始菌株降低了3倍, 酒精得率提高了5倍。首次通过实验验证了热带假丝酵母利用木糖产乙醇的可行性, 这对研究酵母利用秸秆、麦糠、谷壳等纤维质农业废弃物生产燃料乙醇具有重要启示。     

代谢工程改善野生酵母利用木糖产乙醇的性能

从256个自然样品中筛选得到1株可高效转化D-木糖的酵母。通过生理生化和分子生物学方法鉴定, 证实该菌株是属于Candida tropicalis。以该酵母为研究对象, 增加木糖醇脱氢酶表达量, 通过改变代谢流以达到提高酒精产率的目的。以pXY212-XYL2质粒为基础载体, 构建了含有潮霉素抗性的pYX212-XYL2-Hygro, 电击转化进入野生型C. tropicalis, 潮霉素抗性筛选, 得到含高拷贝木糖醇脱氢酶基因的重组菌株C. tropicalis XYL2-7。重组菌的比酶活达到0.5 u/mg protein, 比原始菌株提高了3倍。实验表明, 重组菌木糖醇得率比原始菌株降低了3倍, 酒精得率提高了5倍。首次通过实验验证了热带假丝酵母利用木糖产乙醇的可行性, 这对研究酵母利用秸秆、麦糠、谷壳等纤维质农业废弃物生产燃料乙醇具有重要启示。     

Purification and properties of the proteinase inhibitors from Erythrina caffra (coast erythrina) seed

• 1.1. Four proteinase inhibitors (DE-1 to DE-4) were purified from Erythrinu caffra seed by gel filtration on Sephadex G-50 followed by ion-exchange chromatography involving DEAE-cellulose and DEAE-sepharose.
• 2.2. They comprise 164–166 amino acid residues (mol. wt 18,100) including 4 half-cystine residues and resemble the Kunitz-type proteinase inhibitors.
• 3.3. The N-terminal primary structure of DE-3 revealed also homology with those of the Kunitz-type inhibitors. For DE-1, DE-2 and DE-4 no free N-terminal amino acid was found.
• 4.4. DE-1 contains a potent inhibitor for both porcine trypsin and bovine α-chymotrypsin. Whereas DE-2 inhibits a-chymotrypsin strongly and has practically no action on trypsin, DE-3 inhibits both trypsin and a-chymotrypsin strongly. DE-4 is a potent inhibitor for trypsin but it binds a-chymotrypsin only weakly.

     

Binding of the Kunitz-type trypsin inhibitor DE-3 from Erythrina caffra seeds to serine proteinases: a comparative study.

The effect of pH and temperature on kinetic and thermodynamic parameters (i.e., k(on),k(off),Ka,delta G0, delta H0 and delta S0 values) for the binding of the Kunitz-type trypsin inhibitor DE-3 from Erythrina caffra seeds (ETI) to bovine beta-trypsin, bovine alpha-chymotrypsin, the human tissue plasminogen activator, human alpha-, beta- and gamma-thrombin, as well as the M(r) 33,000 and M(r) 54,000 species of the human urinary plasminogen activator (also named urokinase) has been investigated. At pH 8.0 and 21.0 degrees C: (i) values of the second-order rate constant (K(on)) for the proteinase:ETI complex formation vary between 8.7 x 10(5) and 1.4 x 10(7)/M/s; (ii) values of the dissociation rate constant (k(off)) for the proteinase: ETI complex destabilization range from 3.7 x 10(-5) to 1.4 x 10(-1)/s; and (iii) values of the association equilibrium constant (Ka) for the proteinase:ETI complexation change from < 1.0 x 10(4) to 3.8 x 10(11)/M. Thus, differences in k(off) values account mostly for the large changes in Ka values for ETI binding. The affinity of ETI for the serine proteinases considered can be arranged as follows: bovine beta-trypsin > human tissue plasminogen activator > bovine alpha-chymotrypsin > human alpha-, beta- and gamma-thrombin approximately M(r) 33,000 and M(r) 54,000 species of the human urinary plasminogen activator. Moreover, the serine proteinase:ETI complex formation is an endothermic, entropy-driven, process.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lipid Accumulation during Canola Seed Germination in Response to Cinnamic Acid Derivatives

The objective of this research was to investigate how ferulic and p-coumaric acids affect lipid and fatty acid composition during canola (Brassica napus L.) seed germination. Data showed that both compounds increased total lipid and fatty acid contents in the cotyledons during germination. The largest accumulation in lipids occurred at 1.0 mM p-coumaric acid with an increase in all unsaturated fatty acids. The results suggest that allelochemicals interfere in canola seed germination by reducing lipid mobilization.     

Nitrate and nitrate reductase in Erythrina caffra seeds: Enhancement of induction by anoxia and possible role in germination

The nitrate level in seed embryonic axes of Erythrina caffra Thunb. which is capable of anaerobic germination, was about 2.5 times higher than in seed axes of Pisum sativum L. a species incapable of anaerobic germination. Nitrate levels in E. caffra seeds decreased during germination and this was not due to leaching. Both NADH- and NADPH-dependent nitrate reductase (NAR) activities increased during germination. The increase was prevented by cycloheximide. The activity of NADH-NAR (EC 1.6.6.1) was higher than that of NADPH-NAR (EC 1.6.6.3). Both NAR activities were higher in anoxia than in air during germination. The NAR activities in Pisum seeds were very much lower than in Erythrina seeds. Anoxia (N₂ or argon) enhanced the induction of NAR by KNO₃ in germinated E. caffra axes. The NADH- and NADPH-NAR activities were induced to equally high levels by KNO₃ under anoxia. The enhancement was depressed by cycloheximide. It is concluded that nitrate and NAR activity may play a role in the anaerobic germination of E. caffra seeds.Abbreviation NAR nitrate reductase Financial support was obtained from the University of the Orange Free State and the Foundation for Research Development     

Peanut Resistance Gene Expression in Response to Aspergillus flavus Infection During Seed Germination

Aspergillus flavus produces potent mutagenic and carcinogenic polyketide‐derived secondary metabolites known as aflatoxins. Development of host plant resistance in peanut and other crops is the most environmentally friendly and cost‐effective method to eliminate the serious problem of aflatoxin contamination in grains. To confirm that putative peanut genes identified in a previous microarray study were involved in peanut resistance to A. flavus infection, 14 genes were selected for further investigation through real‐time PCR. The results revealed diverse patterns of gene expression during seed germination after A. flavus inoculation. Based on the expression levels and the relative‐expression patterns over a 7‐day period, the 14 host genes could be classified into six different groups belonging to three main biochemical and genetic defence processes of lipid metabolism, oxidative signalling and cell‐wall synthesis during counter‐attack. A network of gene expression patterns was activated in sequential order in response to A. flavus invasion in both resistant and susceptible peanut lines during seed germination. Understanding gene expression patterns in peanut will be useful to breeders and other scientists interested in incorporating genetic resources of resistance against A. flavus into peanut germplasm and/or commercial cultivars via conventional and/or molecular methods.     

Occurrence of an Inhibitor of Tissue-Type Plasminogen Activator in Seeds and in Vitro Cultures of Erythrina caffra Thunb

The level of an inhibitor of tissue-type plasminogen activator (t-PA) increased slowly during the early developmental stage of seeds of Erythrina caffra Thunb. Thereafter, the inhibitor increased exponentially until the seeds reached maturity. At maturity, the t-PA inhibitor levels in the cotyledons were 38 times higher than the levels at the onset of seed development. The t-PA inhibitor accumulated at a faster rate than the storage proteins, which reached a concentration 15 times higher than the protein concentration at the onset of seed development. During the imbibition and germination process, the t-PA inhibitor decreased gradually. The inhibitor kept on decreasing during the growth of the seedlings until the 10th day after imbibition, when it leveled off at 4.1% of that of the initial inhibitor concentration. The inhibitor remained at this level until the cotyledons were shed at day 22. The total protein in the cotyledons decreased at a slower rate than the inhibitor and reached a minimum concentration at day 20 of 3.6% of the initial protein concentration in the cotyledons. Callus cultures of root, shoot, leaf, and cotyledonary tissue was established and maintained on Murashige-Skoog medium supplemented with 3% sucrose, 10 micromolar benzyladenine, and 5 micromolar 2,4-dichlorophenoxyacetic acid. A shoot cell suspension culture was established on Murashige-Skoog medium supplemented with 3% sucrose, 1 micromolar benzyladenine, and 0.5 micromolar 2,4-dichlorophenoxyacetic acid (pH 5.7) and shaken at 60 revolutions per minute. The level of t-PA inhibitor in root, shoot, leaf, and cotyledonary callus was substantially lower than in the corresponding intact tissue. The t-PA inhibitor levels in the linear growth phase was higher than in the lag or stationary growth phases of the cell suspension culture. A hydrolysate of the cell walls of tomato and E. caffra Thunb, as well as polyamines and organic acids, did not increase the concentration of t-PA inhibitor in suspension cultures or intact leaf tissue of E. caffra. The t-PA inhibitor levels of suspension cultures were increased by Na₂SO₄ but not by I-cysteine in the nutrient medium.     

Action of Respiratory Inhibitors on Seed Germination and Oxygen Uptake in Avena fatua L.

The effects of sodium azide, potassium cyanide (cytochrome oxidaseinhibitors), and salicylhydroxamic acid (SHAM; an alternativerespiration inhibitor) on germination and respiration of Avenafatua L. seeds were studied. Azide and cyanide released seeddormancy at similar concentrations and treatment durations.Cyanide, however, stimulated germination of seeds with littleafter-ripening, whereas azide had no effect under similar conditionsunless the seeds were after-ripened for several months; theduration of after-ripening required for seeds to respond toazide varied with seed batch. There was also a greater lag priorto germination in the case of azide, compared to cyanide treatedseeds. SHAM inhibited the stimulation of germination and respirationby azide, but not by cyanide. Furthermore, respiration induced by azide or cyanide could notbe inhibited by the subsequent application of SHAM. These findingssuggest that the respiration stimulated by azide and cyanideis not alternative (SHAM-sensitive) and, therefore, this respiratorypathway cannot be involved in the stimulation of germinationby cytochrome oxidase inhibitors. While embryos excised fromcontrol, azide or cyanide pretreated seeds had the capacityto perform alternative respiration, the actual contributionof this pathway was negligible. A large proportion of respirationof embryos excised from azide or cyanide pretreated seeds wasresidual, i.e. insensitive to both SHAM and cyanide. Alternative respiration, azide, cyanide, dormancy, salicylhydroxamic acid, wild oats     

Mechanism of Fermentation Conversion from Polyalcohol Fermentation to Ethanol Fermentation by Pichia miso

A possible mechanism of fermentation conversion is described from polyalcohol fermentation to ethanol fermentation by Pichia miso. Little alcohol dehydrogenase activity was found in polyalcohol-producing cells, whereas higher enzyme activity was induced by ethanol-producing cells. The fermentation conversion may be caused by the different levels of alcohol dehydrogenase activity between polyalcohol- and ethanol-producing cells. It was also shown that yeast growth was inhibited and that yeast cells were lysed by ethanol (at 6g/100ml) that accumulated in 24 hr.     

拟南芥AtR8 lncRNA对盐胁迫响应及其对种子萌发的调节作用

长链非编码RNA (lncRNA)是一类长度大于200个核苷酸且不编码蛋白质的非编码RNA, 主要由RNA聚合酶II转录生成, 大量存在于生物体内并具有多种生物学功能。AtR8 lncRNA是拟南芥(Arabidopsis thaliana)中RNA聚合酶III转录的长链非编码RNA。前期研究发现, 水杨酸(SA)处理诱导萌发种子中AtR8 lncRNA的表达, AtR8 lncRNA缺失抑制SA胁迫下的种子萌发。进一步研究发现, AtR8 lncRNA转录区域内存在保守的盐胁迫响应元件(TCTTCTTCTTTA); NaCl处理抑制萌发种子中AtR8 lncRNA的表达; 与野生型相比, 高浓度NaCl处理明显抑制了atr8 (AtR8 lncRNA部分缺失型拟南芥)种子萌发。研究结果表明, AtR8 lncRNA在拟南芥种子萌发期的盐胁迫中起重要作用。     

Phytochrome and Seed Germination: VI. Phytochrome and Temperature Interaction in the Control of Cucumber Seed Germination

Phytochrome control of cucumber seed germination is temperature-dependent. A prolonged exposure to radiation from broad spectrum far red sources (Pfr/P = 0.05 to 0.07) prevents germination at temperatures below 20 C. Above 20 C there is no inhibition and it appears as if there is an escape from phytochrome control. However, radiation from a monochromatic, narrow band 730 nanometer source (Pfr/P < 0.02) inhibits germination at temperatures above 20 C. This result supports the idea that, even at high temperatures, Pfr is responsible for the activation of germination. After 4 days of exposure to far red, a short red irradiation is quite effective in promoting germination if temperatures during the dark incubation periods are maintained below 20 C; red becomes effective at temperatures above 20 C. Promotion of germination will take place at a temperature of 25 C or higher without red irradiation. Again, we have an apparent escape from phytochrome control at high temperatures. However, if higher temperatures are used for only short periods, 2 to 6 hours, in combination with short red irradiation, one can demonstrate that activation of germination at high temperatures is still dependent on phytochrome. Phytochrome is probably destroyed during prolonged exposure to far red. Thus, the subsequent short red irradiation establishes levels of Pfr which may not be sufficient to promote germination at low temperatures but are probably adequate at high temperatures.     

The Release of Nitrite from Barley Roots in Response to Metabolic Inhibitors, Uncoupling Agents, and Anoxia

When excised sterile barley roots, from plants which had beengrown in the presence of nitrate, were placed under low oxygentensions, nitrite was released into the external solution. Themaximum leakage of nitrite occurred under completely anaerobicconditions. Nitrite was also released from barley roots underaerobic conditions when uncouplers of oxidative phosphorylation(DNP, CCCP¹, pentachlorophenol) or certain simple organic acidswere supplied. Inhibitors of the Krebs cycle, or of respiratoryelectron transport, were much less effective in causing theloss of nitrite, possibly because these compounds did not ingeneral inhibit root respiration severely. Nitrite release, in response to any of the above treatments,was accompanied by an accumulation of nitrite within the tissue.It was concluded that an increase in membrane permeability,and a decrease in ATP synthesis, were contributory causes ofthis phenomenon, although neither could explain the experimentalobservations completely. There was however no evidence thatthe pentose phosphate pathway, which is regarded as the sourceof reducing power for nitrite reduction, was inhibited underconditions which favoured nitrite release.