Partial decline of Arachis hypogaea L. photosynthesis triggered by drought stress

Photosynthetic capacity (PC) of three peanut cultivars (Arachis hypogaea L. cvs. 57-422, 73-30, and GC 8-35) decreased during drought stress (decline in relative water content from ca. 95 to 70 %) and recovered two days after rewatering. Mild water stress was not limiting for the total ribulose-1,5-bisphosphate carboxylase/oxygenase activity, since this enzyme activity increased under drought. Photosystem (PS) 2 and PS1 (the latter only in cv. GC 8-35) electron transport activities decreased under drought. The ratio of the variable to maximal chlorophyll fluorescence (Fv/Fm) decreased mainly in the cv. GC 8-35. All cultivars showed decreases in photochemical quenching (qP) and quantum yield of PS2 electron transport (Φe). Increase of basal fluorescence (F0) was observed in the cvs. 73-30 and GC 8-35, while the cv 57-422 showed a decrease. After rewatering a sharp increase was observed in the majority of the parameters. Thus under the present stress conditions, the cv GC 8-35 was the most affected for all the parameters under study. The cv. 57-422 showed a higher degree of tolerance being gradually affected in photosynthetic capacity (PC) in contrast to the two other cvs. which showed a sharp decrease in PC at the beginning of the drought cycle. This revised version was published online in September 2006 with corrections to the Cover Date.     

基因重组大肠杆菌表达血红素加氧酶-1及培养条件优化

【背景】血红素加氧酶-1 (HO-1)具有抗氧化应激、抗凋亡和抗纤维化等多种生理效应,有望成为一种新型药物应用于临床疾病的治疗。【目的】构建表达HO-1的基因重组大肠杆菌(Escherichiacoli),并优化其表达培养条件,实现HO-1高产率的表达。【方法】PCR法克隆集胞藻(Synechocystissp.)PCC6803的HO-1基因(ho1),构建重组质粒pET-28a-ho1,转化大肠杆菌BL21(DE3)菌株,单因素实验优化表达培养基的种类、诱导剂添加时间、诱导培养时间、诱导剂浓度和诱导培养温度。【结果】构建了表达HO-1的基因重组大肠杆菌BL21(DE3)/pET-28a-ho1菌株,用甘油(GY)培养基培养至菌体浓度OD_(600)约为0.8时,加入终浓度为0.1 mmol/L的IPTG诱导,30°C诱导培养6 h,HO-1的表达量最高,Ni-NTA柱分离纯化得到的HO-1收率占细胞总蛋白的10.9%。【结论】获得了可溶性表达HO-1的基因重组大肠杆菌及其较佳的培养条件,为进一步研究集胞藻来源的HO-1的酶学性质和应用奠定了基础。     

Alternate paths of evolution for the photosynthetic gene rbcL in four nonphotosynthetic species of Orobanche

We have determined the nucleotide sequence for the Rubisco large subunit from four holoparasitic species of Orobanche. Intact open reading frames are present in two species (O. corymbosa and O. fasciculata), whereas the remaining species (O. cernua and O. ramosa) have rbcL pseudogenes. Sequences for rbcL 5'-UTRs from species of Orobanche have few changes in the promoter and ribosome binding sites compared to photosynthetic higher plants. Comparison of rbcL 3'-UTR sequences for Nicotiana, Ipomoea, Cuscuta, and Orobanche reveal that nucleotide sequences from parasitic plants have regions capable of forming stem-loop structures, but 56–69 nt are deleted upstream of the stem-loop in the parasitic plants compared to their photosynthetic relatives. Although rbcL pseudogenes of O. cernua and O ramosa have many large and small deletions, few indels are shared in common, implying that their common ancestor probably had an intact rbcL reading frame. Intact rbcL reading frames in O. corymbosa and O. fasciculata retain a bias of synonymous over nonsynonymous substitutions and deduced protein sequences are consistent with potentially functional Rubisco large subunit proteins. A conservative model of random substitution processes in pseudogene sequences estimates that the probability is low (P<0.028) that these sequences would retain an open reading frame by chance. Species of Orobanche have either had recent photosynthetic ancestors, implying multiple independent losses of photosynthesis in this genus, or the rbcL gene may serve an unknown function in some nonphotosynthetic plants.     

The homologous tryptophan critical for cytochrome c peroxidase function is not essential for ascorbate peroxidase activity

The crystal structures of ascorbate peroxidase (APX) and cytochrome c peroxidase (CCP) show that the active site structures are nearly identical. Both enzymes contain a His-Asp-Trp catalytic triad in the proximal pocket. The proximal Asp residue hydrogen bonds with both the His proximal heme ligand and the indole ring nitrogen of the proximal Trp. The Trp is stacked parallel to and in contact with the proximal His ligand. This Trp is known to be the site of free radical formation in CCP compound I and also is essential for activity. However, APX forms a porphyrin radical and not a Trp-centered radical, even though the His-Asp-Trp triad structure is the same in both peroxidases. We found that conversion of the proximal Trp to Phe has no effect on APX enzyme activity and that the mutant crystal structure shows that changes in the structure are confined to the site of mutation. This indicates that the paths of electron transfer in CCP and APX are distinctly different. The Trp-to-Phe mutant does alter the stability of the APX compound I porphyrin radical, by a factor of two. Electrostatic calculations and modeling studies show that a potassium cation located about 8?Å from the proximal Trp in APX, but absent in CCP, makes a significant contribution to the stability of a cation Trp radical. This underscores the importance of long-range electrostatic effects in enzyme catalyzed reactions.     

Effects of ultraviolet radiation on photosynthesis and related enzyme reactions of marine macroalgae

 Changes in physiological parameters related to photosynthesis were studied in five macroalgal species from Spitsbergen (Monostroma arcticum, Laminaria solidungula, Alaria esculenta, Palmaria palmata, Phycodrys rubens) during a 72-h exposure to UV radiation. Maximal quantum yield of photochemistry (Fv/Fm) and maximal electron transport rate (ETRmax) were measured with a pulse-amplitude-modulated fluorometer; the activity of the Calvin cycle enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) were estimated using a photometric test. Proteins of crude extracts were separated by SDS gel electrophoresis and changes in cellular concentrations of Rubisco were determined. Moreover, the concentration of chlorophyll a (Chl a), and protein content, were measured photometrically. In all species, Chl a content, maximal quantum yield as well as ETRmax decreased during the UV treatment. Changes in ETRmax were related to the changes in the overall activity of Rubisco. Analysis of SDS gels showed that in P. rubens, L. solidungula, M. arcticum and A. esculenta decreasing Rubisco activity partly resulted from a degradation of the enzyme. However, in A. esculenta, the formation of a high-molecular-weight polypeptide was observed. In all species, the activity of Rubisco was more strongly impaired than that of G3PDH. Exposure to UV resulted in loss of total protein only in the deepwater species L. solidungula and P. rubens. The different sensitivities to UV exposure of the species tested reflect their zonation pattern in the field. Received: 4 October 1999 / Accepted: 15 February 2000     

Fractionation of erytroblasts with affinitymediated modifications of their electrical properties using counter-current distribution

Enterally administered, heme is a good source of iron in humans and other animals, but the metabolism of heme by enterocytes has not been fully characterized. Caco-2 cells in culture provide a useful model for studying cells that resemble small intestinal epithelium, both morphologically and functionally. In this paper we show that heme oxygenase, the rate-controlling enzyme of heme catabolism, is present in abundance in Caco-2 cells, and that levels of its mRNA and activity can be increased by exposure of the cells to heme or metal ions (cadmium, cobalt). Caco-2 cells also contain biliverdin reductase activity which, in the basal state, is similar to that of heme oxygenase (approximately 40 pmole of product per mg protein per minute); however, when heme oxygenase is induced, biliverdin reductase may become rate-limiting for bilirubin production.Abbreviations BVR biliverdin reductase - DMEM Dulbecco's modified Eagles medium - DMSO dimethyl sulfoxide - HO heme oxygenase - 1xSSC a solution of 0.015 M sodium citrate/0.15 sodium chloride     

拟南芥色氨酸与吲哚乙酸生物合成的研究进展

拟南芥色氨酸生物合成途径的研究已逐渐成为植物分子生物学家了解植物基因结构和表达调控最主要的模式系统之一。到目前为止,编码拟南芥色氨酸合成途径的七种酶蛋白的基因已经全部被克隆,并进行了不同程度的分子生物学研究。长期以来,色氨酸一直被认为是植物生长素吲哚乙酸（ＩＡＡ）生物合成（从头合成）的前体物,但近年来人们发现生长素合成的非色氨酸途径可能是其在植物中生物合成的主要途径。植物在不同的发育阶段可能采用不同的方式合成ＩＡＡ。     

Aromatic dioxygenases: molecular biocatalysis and applications

Aromatic dioxygenases have been found to catalyse single and tandem oxidation reactions of conjugated polyenes. Rational selection and design of dioxygenases, allied to substrate shape, size and substitution pattern, has been used to control regiochemistry and stereochemistry during the oxygenation process. The resulting enantiopure bioproducts have been increasingly utilised as precursors for new and alternative routes in chiral synthesis.     

Sleep quality and duration following evening intake of alpha-lactalbumin: a pilot study#

Tryptophan loading enhances sleep quality by increasing the ratio of plasma tryptophan to large neutral amino acids (TRP:LNAA) and consequently synthesis and availability of serotonin in the brain. Alpha-lactalbulmin (A-LAC) is rich in tryptophan and has the highest TRP:LNAA of all protein sources. This pilot study investigated the effect of an evening intake of A-LAC on objective and subjective sleep measures in male subjects without sleep complaints. Ten healthy male university students (aged: 26.9 ± 5.3 years; BMI: 21.7 ± 1.9 kg.m^?2) participated in a double-blind, randomized, and placebo-controlled crossover counter-balanced study. Objective (actigraphy) and subjective (sleep log) sleep measures were recorded for two nights after a standardized evening meal supplemented with either A-LAC (20 g) or a placebo of sodium caseinate (20 g) one hour before bedtime. Evening A-LAC intake resulted in increased objective and subjective total sleep time by 12.8% (p = 0.037) and 10.8% (p = 0.013), respectively, compared to placebo. Objective sleep efficiency increased by 7.0% (p = 0.028) following A-LAC with no significant effects for other sleep indices. This pilot study demonstrates the efficacy of evening A-LAC intake on sleep quality in young healthy adults, however further large-scale studies are warranted to confirm the benefit.