Free amino acid contents of the gametophytic blades from the green mutant conchocelis and the heterozygous conchocelis in Porphyra yezoensis Ueda (Bangiales, Rhodophyta)

Free amino acid contents in green mutant(G-1) blades and sectored F₁gametophytic blades with green andwild-type portions, which were developedfrom heterozygous conchocelis obtained by across between the wild type (0110) and thegreen mutant (G-1) of Porphyrayezoensis, were compared with those of thewild-type blades in laboratory culture. The contents of the major four free aminoacids (aspartic acid, glutamic acid,alanine and taurine) as well as those ofthe total free amino acids were highest inthe green mutant blades, intermediate inthe F₁ gametophytic blades, and lowestin the wild-type blades. A similar trendwas obtained in the blades developed frommonospores of the F₁ gametophyticblades. In addition, the green-typesectors also had a higher content of thefour major free amino acids and total freeamino acids compared with the wild-typesectors in the F₁ blades cultivated ata nori farm. The green mutant ischaracterized by higher contents of thefour major free amino acids compared withthe wild type, which has a higher growthrate. Hence, it is considered that thesectored F₁ gametophytic bladesproduced from the heterozygous conchocelishave both parental advantages (high freeamino acid contents and high growth rate)and compensate for both parentaldisadvantages. This seems to be one of thepossible ways of genetic improvement inregards to the taste of nori and stableproduction in Porphyra cultivation.     

Effects of NaCl-salinity on amino acid and carbohydrate contents of Phragmites australis

The effects of NaCl-salinity on growth, free amino acid and sugar content and composition were assayed in roots, rhizomes and leaves of Phragmites australis (Cav.) Trin. ex Steud. Juvenile plants produced from freshwater clones, were cultured under greenhouse hydroponic conditions for 21 days. Relative growth rates were highest at a salinity level of 0 and 1.5‰, respectively, but decreased significantly at 10‰. All plants cultured at 35‰ salinity died. The osmolality in rhizomes and leaves increased with salinity. The total contents of free amino acids were highest in rhizomes>leaves>roots. In rhizomes, the amino acid content increased significantly up to four-fold from 0 to 10‰ salinity. This increase was caused by up to 200-fold increase of proline and 11-fold increase of glutamine at 10‰, whilst the share of asparagine and glutamate decreased. Leaves showed a similar response to salinity with increasing amino acid contents, and shares of proline and glutamine whereas roots did not react significantly. The contents of sucrose, glucose and fructose were highest in leaves>rhizomes>roots. In rhizomes of all three clones, the sugar contents increased up to 3.5-fold from 1.5 to 10‰ salinity level, but were lower at 1.5‰ versus the control (0‰). Sugar contents were lowest (roots) and highest (leaves) at 1.5‰ salinity. The sugar composition did not vary significantly except for leaves where the fraction of sucrose decreased with increasing salinity level at all three clones from 89.1 to 61.7% of total dissolved sugar (pooled data). The importance of free amino acids and sugars as osmolytes was similar in rhizomes and leaves (13–15% of total osmolality at 10‰). In rhizomes, free amino acids were more important as osmolyte than sugars, while the opposite was true for leaves. Proline contributed up to 2.7% to total osmolality. It is hypothesised that a strong proline accumulation indicates the exceeding of a critical salinity level.     

转CiCHS基因拟南芥的黄酮代谢及抗氧化能力分析

该研究克隆了中间锦鸡儿的查尔酮合成酶基因(CiCHS)并转入野生型拟南芥和tt4突变体,用qRT-PCR检测了转基因拟南芥中内源AtCHS基因的表达量,用分光光度法分析了转基因拟南芥的总黄酮、丙二醛含量及DPPH自由基清除能力,用HPLC法检测了转基因拟南芥的柚皮苷含量。结果显示:(1)转基因拟南芥中,内源AtCHS基因的表达量约为野生型的十分之一,总黄酮含量明显高于野生型;HPLC测得转基因株系中柚皮苷含量高于野生型;紫外照射处理前后转基因拟南芥中丙二醛积累量明显少于野生型。(2)转基因株系提取物对DPPH自由基清除能力显著高于野生型。(3)CiCHS基因互补拟南芥tt4突变体,转基因株系的种皮呈现浅棕色。研究表明,中间锦鸡儿CiCHS基因异源表达后生成了柚皮苷,使转基因植物的抗氧化性增强,部分恢复了tt4突变体的种皮颜色。     

Seasonal patterns of nucleic acid concentrations and amino acid profiles of Parapenaeus longirostris (Crustacea, Decapoda): relation to growth and nutritional condition

The present work describes the seasonal changes in nucleic acid concentrations and amino acid profiles in the muscle of juvenile Parapenaeus longirostris and their relation to growth and nutritional condition. RNA content varied significantly between seasons, being the highest values attained in spring and the lowest in winter (p < 0.05). Similar results were obtained with RNA:protein and RNA:DNA ratios. In respect to total amino acid content (TAA), a significant increase from winter to spring was observed (p < 0.05) and the major essential amino acids (EAA) were arginine, histidine and leucine. Within non-essential amino acids (NEAA) glutamic acid, aspartic acid, glycine and proline were dominant. From winter to spring, a significant variation in NEAA content occurred (26.8; p < 0.05), mainly due to the significant increase of glutamic acid (79.1) and serine (66.7) (p < 0.05). EAA content did not vary significantly between seasons (p > 0.05). In opposition, during this period a significant decrease in the free amino acid content (FAA) was observed (p < 0.05); a higher percentage of decrease was attained in free non-essential (FNEAA – 42.9) in comparison to free essential amino acids (FEAA – 40.2). The significant increase in RNA and TAA contents from winter to spring may be related with protein synthesis. On the other hand, the lowest values obtained in winter may be due to a reduction in feeding activity; in this period the muscle protein must be progressively hydrolysed, which is evident with the higher FAA content. The liberated amino acids enter FAA pool and become available for energy production. In conclusion, it was evident that the seasonal cycle in activities such as feeding and growth with nucleic acids and amino acid analyses was noticed.     

走马胎叶片营养成分分析及栽培年限差异比较

走马胎为西南地区常用中药材,被广泛用于食物蒸炖、药浴等日常保健。近年来走马胎叶片被开发成代泡茶,有效拓宽了应用途径。为了明确走马胎叶片营养价值,该研究采用国标方法分别对不同栽培年限走马胎叶片的矿质元素、一般营养、氨基酸及活性成分进行了分析和评价。结果表明：(1)走马胎叶片含有非常丰富的矿质营养,并符合高钾低钠型保健食品特征。1年生植株叶片矿质元素含量最高,其中磷(P)、铁(Fe)、锰(Mn)、锌(Zn)显著高于2～4年生植株(P<0.05)。(2)走马胎叶片的一般营养和氨基酸含量较高,且随栽培年限不同存在明显差异;4年生植株叶片维生素C、总糖和氨基酸含量显著高于1～3年植株(P<0.05)。(3)走马胎叶片含有丰富的酚类和皂苷类物质,黄酮类物质含量较低,其中1年生总皂苷和4年生总酚含量均显著高于其他栽培年限的植株叶片(P<0.05)。综上研究表明,与常用果蔬及茶叶相比,走马胎叶片营养价值较高,其中1年生的植株叶片中矿质营养和皂苷类物质的总含量最高,4年生的植株叶片中一般营养(除脂肪外)、氨基酸及酚类物质的总含量最高,具有较好的开发利用前景。     

Increasing lysine levels in pigeonpea (<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp) seeds through genetic engineering

In higher plants the essential amino acids lysine, threonine, methionine and isoleucine are synthesised through a branched pathway starting from aspartate. The key enzyme of lysine biosynthesis in this pathway—dihydrodipicolinate synthase (DHDPS)—is feedback-inhibited by lysine. The dhdps-r1 gene from a mutant Nicotiana sylvestris, which encodes a DHDPS enzyme insensitive to feedback inhibition, was used to improve the lysine content in pigeonpea seeds. The dhdps-r1 coding region driven by a phaseolin or an Arabidopsis 2S2 promoter was successfully overexpressed in the seeds of pigeonpea by using Agrobacterium transformation and particle bombardment. In 11 lines analysed, a 2- to 6-fold enhanced DHDPS activity in immature seeds at a late stage of maturation was found in comparison to wild type. The overexpression of dhdps-r1 led to an enhanced content of free lysine in the seeds of pigeonpea from 1.6 to 8.5 times compared with wild type. However, this was not reflected in an increase in total seed lysine content. This might be explained by a temporal discrepancy between maximal expression of dhdps-r1 and the rate of amino acid incorporation into storage proteins. Assays of the lysine degradative enzyme lysine-ketoglutarate reductase in these seeds showed no co-ordinated regulation of lysine biosynthesis and catabolism during seed maturation. All transgenic plants were fertile and produced morphologically normal seeds.     

Mulberry Leaf Metabolism under High Temperature Stress

Effects of high temperature on the activity of photosynthetic enzymes and leaf proteins were studied in mulberry (Morus alba L. cv. BC2-59). A series of experiments were conducted at regular intervals (120, 240 and 360 min) to characterize changes in activities of ribulose-1,5-bisphosphate carboxylase (RuBPC) and sucrose phosphate synthase (SPS), photosystem 2 (PS 2) activity, chlorophyll (Chl), carotenoid (Car), starch, sucrose (Suc), amino acid, free proline, protein and nucleic acid contents in leaves under high temperature (40 °C) treatments. High temperature markedly reduced the activities of RuBPC and SPS in leaf extracts. Chl content and PS 2 activity in isolated chloroplasts were also affected by high temperature, particularly over 360 min treatment. Increased leaf temperature affected sugar metabolism through reductions in leaf starch content and sucrose-starch balance. While total soluble protein content decreased under heat, total amino acid content increased. Proline accumulation (1.5-fold) was noticed in high temperature-stressed leaves. A reduction in the contents of foliar nitrogen and nucleic acids (DNA and RNA) was also noticed. SDS-PAGE protein profile showed few additional proteins (68 and 85 kDa) in mulberry plants under heat stress compared to control plants. Our results clearly suggest that mulberry plants are very sensitive to high temperature with particular reference to the photosynthetic carbon metabolism.     

湖南道地白芷氨基酸含量与组成分析

白芷是我国常用中药材的一种,关于白芷镇痛的有效成分,主要认为是异欧前胡素、欧前胡素等香豆素类具有解痉、镇痛等作用的挥发油类化学成分,但白芷镇痛的成分及其机理尚不十分清楚,是否有其他物质也具有一定的药理作用也不得而知,比如说某些药效性氨基酸。该研究采用自动氨基酸分析仪测定了湖南道地茶陵白芷的蛋白质类氨基酸和游离氨基酸的含量,并分析其氨基酸组成。结果表明:总氨基酸检出除Asn以外的其他16种蛋白质氨基酸,含量最高的Arg占总氨基酸的31.21%,接近1/3;必需氨基酸总量达27.01%,其中含量最高的是Leu,占总必需氨基酸的24.14%;药效氨基酸比例较高,在酸水解产物总氨基酸和游离氨基酸中分别达到73.89%和85.78%,其中Arg的含量最高,达1.383g·100g-1,占比为42.24%,游离氨基酸中γ-氨基丁酸、鸟氨酸含量也较高。此外,还含有少量的高半胱氨酸和鹅肌肽等游离非蛋白质氨基酸和短肽;必需氨基酸的组成接近WHO/FAO的建议摄入值,但Met+Cys的RC值最小,为第一限制性氨基酸。这些药效性氨基酸和游离氨基酸可能是茶陵白芷具有良好药效的一个因素,而且茶陵白芷的氨基酸组成和配比较合理、符合人体需要,在强化含硫氨基酸的基础上具有开发成为新型药食同源保健性食品的潜力。研究结果可为进一步研究白芷的药理作用提供依据。     

Changes in Mulberry Leaf Metabolism in Response to Water Stress

A series of experiments were conducted to characterize the water stress-induced changes in the activities of RuBP carboxylase (RuBPCO) and sucrose phosphate synthase (SPS), photosystem 2 activity, and contents of chlorophylls, carotenoids, starch, sucrose, amino acids, free proline, proteins and nucleic acids in mulberry (Morus alba L. cv. K-2) leaves. Water stress progressively reduced the activities of RuBPCO and SPS in the leaf extracts, the chlorophyll content, and PS2 activity in isolated chloroplasts. Plants exposed to drought showed lower content of starch and sucrose but higher total sugar content than control plants. While the soluble protein content decreased under water stress, the amino acid content increased. Proline accumulation (2.5-fold) was noticed in stressed leaves. A reduction in the contents of DNA and RNA was observed. Reduced nitrogen content was associated with the reduction in nitrate reductase activity. SDS-PAGE protein profile showed few additional proteins (78 and 92 kDa) in the water stressed plants compared to control plants.     

栽培与野生化血丹不同部位中两种化学成分含量分析

为探讨并分析栽培与野生化血丹植株中不同部位中两种化学成分的含量差异,该研究采用超声法提取、高效液相色谱法(HPLC)测定栽培与野生化血丹根、茎、叶、花、混合样等部位中桃叶珊瑚苷和梓醇的含量,并进行比较。结果表明:(1)桃叶珊瑚苷在栽培与野生化血丹植株内均有分布,含量均以根中最高,其在栽培与野生化血丹植株内的含量表现分别为根叶混合样茎花、根混合样茎花叶,栽培化血丹不同部位中桃叶珊瑚苷的含量均高于野生化血丹。(2)梓醇在栽培化血丹的茎中未检出,在栽培与野生化血丹其他部位均有分布,含量均以叶中最高,其在栽培与野生化血丹植株内的含量分别表现为叶花混合样根、叶混合样茎花根,野生化血丹不同部位中梓醇的含量均高于栽培化血丹。(3)桃叶珊瑚苷和梓醇在栽培和野生化血丹植株不同部位中的含量均存在显著差异(P0.05),栽培与野生同一部位间总体上无显著差异,为该濒危药用植物资源药用部位选择和合理开发利用提供实验参考。     

Lipid Composition of Citrus Leaves from Plants Resistant and Susceptible to Citrus Bacterial Canker

The contents and composition of lipids in citrus leaves in relation to their general resistance to infection by strains of Xanthomonas campestris pv. citri (Xcc) were determined. The composition and contents of total polar lipids and phospholipids and the degree of fatty acid unsaturation were significantly different between resistant and susceptible species. Leaves from resistant plants had less phospholipids, but more free sterols than those from susceptible plants. The predominant fatty acids in the phospholipids were palmitic (16:0), linoleic (18:2) and α-linolenic acid (18:3). The degree of fatty acid unsaturation was higher in susceptible plants than in resistant plants. Major phospholipids in citrus leaves were phosphatidylchloline (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and phosphatidylinositol (PI). β-Sitosterol, campesterol and lanosterol were major sterols in the leaves of citrus plants with resistant species having a higher ratio of free sterols to total phospholipids than susceptible species. Differences in lipid metabolism may contribute to differences in Xcc-resistance of citrus leaves.     

Alleviation of Changes in Protein Metabolism in NaCl-Stressed Wheat Seedlings by Thiamine

NaCl-stress induced a pronounced suppression in growth of wheat seedlings. The most abundant amino acids (cysteine, arginine, methionine) constituting about 55 % of total free amino acid content in control wheat were reduced in 100 mM NaCl-treated plants. However, valine, isoleucine, aspartic acid and proline accumulated in response to NaCl stress and NaCl-treated wheat seedlings showed 1.6 fold increase in total free amino acids compared to the control. Addition of 2 [micro ]M thiamine alleviated the effects of NaCl on the amino acid composition and the amount of total free amino acids decreased to that in the control. Content of 26 kDa protein increased in NaCl-treated plants, stimulation was more pronounced in roots than in shoots. In contrast, the contents of 13 and 20 kDa proteins decreased. After addition of thiamine, the 24 kDa protein, which disappeared with NaCl treatment, has been initiated again. Moreover, thiamine treatment stimulated the accumulation of the 20 kDa protein.     

Effect of growth temperature and media composition on the fatty acid composition of Bacillus stearothermophilus AN 002

The influence of growth temperature, media composition and cell age on the chemical composition of Bacillus stearothermophilus strain AN 002 has been determined. The total cellular protein decreased and the free amino acid content increased with growth temperature, in both exponential and stationary growth phase. The protein and free amino acid contents of cells were higher in the stationary phase than in the exponential phase, irrespective of growth temperature and media composition. The RNA content was only reduced in cells grown at 55° C. No significant variations were observed in the DNA and carbohydrate contents with respect to growth temperature and cell age. The total lipid and fatty acid compositions on the other hand varied as a function of growth temperature, cell age and media composition. Differences in the relative concentrations of even, odd and branched chain fatty acids were noticed. Novariation was observed in the antiiso and unsaturated fatty acids with respect to growth temperature. The unique variations in the fatty acid composition and total lipids at the growth temperature of 50° C and their variations in the stationary growth phase seem to be characteristic for B. stearothermophilus AN 002.     

Phytochemical analysis of Jatropha curcas L. during different seasons and developmental stages and seedling growth of wheat (Triticum aestivum L) as affected by extracts/leachates of Jatropha curcas L

Jatropha curcas shows invasive characters and is a significant source of many phytochemicals with varying biological activities. Different plant parts of Jatropha curcas L exhibited variation in their phytochemical constituents. Leaves and ovary walls were found to contain higher contents of total phenols, tannins and phytic acid whereas free amino acids were greater in leaves. Young leaves of Jatropha show greater contents of all these metabolites. Further, plants exhibit seasonal differences as leaves collected during summer (May-June) have greater accumulation of total phenols, tannins and free amino acids however, phytic acid was more during rainy season. Leachates and extracts in their higher concentrations adversely affected the germination and growth of wheat seedlings however, lower concentrations were more or less stimulatory. These treatments not only decreased the length, fresh and dry weight of seedlings but also affected the chlorophyll contents and activity of enzymes such as nitrate reductase, aminotransferases in wheat seedlings however, the activity of superoxide dismutase and ascorbate peroxidases increased. Experiments indicate harmful allelopathic effects of Jatropha leachates /extracts on wheat seedlings, hence further experimentation and analysis is recommended before continued plantation of Jatropha particularly on fertile soils. However. Growth of Jatropha plants on saline soils and their potential for accumulating sodium, potassium and chloride are the attributes suggesting the possibility of use of Jatropha plants in improving saline soils.     

Fas- and Mitochondria-Mediated Signaling Pathway Involved in Osteoblast Apoptosis Induced by AlCl<Subscript>3</Subscript>

The concentrations of 13 elements (Al, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, and Zn) were determined in several samples of native (wild) naturally growing and cultivated blueberry fruits. The total metal contents after mineralization were analyzed by inductively coupled plasma optical emission spectrometry. Reliability of the procedure was checked by the analysis of the certified reference materials Mixed Polish Herbs (INGT-MPH-2) and Leaves of Poplar (NCS DC 73350). In the fruits collected in the forest (wild blueberries), higher contents of Ca, Na, and Mg as well as Mn and Zn were observed. Similar levels of Cu, Cr, Fe, and Ni were detected in both wild-growing and cultivated plants. The significantly higher content of Fe and Cd in cultivated blueberries was connected with the content of these metals in soil samples collected from the same places. The metal extraction efficiency by hot water varied widely for the different blueberries (wild or cultivated) as well as their form (fresh or dried).     

Taxonomic variation in total leaf protein amino acid compositions of grasses

Leaves of greenhouse grown grasses had free protein amino acid contents of generally less than 5 % total amino acids, while field collected grasses averaged 14.7 % free protein amino acid contents. Taxonomic patterns are detectable in the total leaf amino acid profiles of grasses from both sources, those of pooids being distinguishable from those of chloridoids and panicoids, and those of danthonioids showing an intermediate pattern. Leaf profiles of Oryza, Stipeae, and Ehrharteae resemble one another, and are more like those of pooids than those of panicoids. Variations in Thr and Leu are apparently associated with differences in photosynthetic pathway. Grass leaves are generally low in total amino acid contents (2.2 ? 1.0 g % fr. wt), with Ile, Val and Met + Cys identified as the limiting essential amino acids. However, the nutritional ‘chemical scores’ of grass leaf proteins are high (75 %, based on the WHO scoring pattern).     

Stress-induced changes in the free amino acid composition in transgenic soybean plants having increased proline content

Following drought stress at supraoptimal temperature the increase in proline (Pro) content in transgenic (T) soybean [Glycine max (L.) Merr. cv. Ibis] plants overexpressing the gene coding for the last enzyme of Pro biosynthesis, L-Δ¹-pyrroline-5-carboxylate reductase, was much greater than in wild type (W) plants (105-fold versus 19-fold after 7 d). Under control conditions arginine accounted for nearly 60 % of the total free amino acid content. After stress treatment the content of Pro was more than 50 % in both T and W genotypes, and at the end of recovery the γ-aminobutyrate content reached 27 and 53 % in the W and T plants, respectively. Without stress treatment there was only a 2-fold difference between T and W in the tyrosine content. However, during the stress period and the subsequent recovery a similar difference was found for many amino acids. The present results indicate that manipulating of the content of a single amino acid influences the whole free amino acid composition in soybean.     

The effect of nitrogen and sulphur on changes in nitrogen fractions of barley plants at various early stages of growth and on yield and amino acid composition of grain

Summary In a pot experiment the effects were estimated of varying nitrogen and sulphur applications on the total and nitrate nitrogen, free amino acid and amide and total amino acid concentrations of barley plants at five early stages of growth.Variations with time in contents of total and nitrate nitrogen differed at low and high nitrogen levels. Concentrations of free amino acids and amides were influenced by light intensity preceding sampling, age of plant and by levels of nitrogen and sulphur. At the fifth harvest (55 days old plants), high nitrogen addition without sulphur caused a fivefold increase in asparagine content and lesser increases in glycine and serine. Alterations in total amino acid contents with time were largely dependent on variations in total nitrogen. Sulphur deficiency, as judged by contents of S-amino acids and amides, became more pronounced with time. A positive interaction was found between effects of nitrogen and sulphur on the yield and cystine and methionine content of grain. Absence of sulphur reduced the cystine content of grain much more than that of methionine, particularly at the high nitrogen level.     

Changes in protein and amino acid content during anther development in fertile and cytoplasmic male sterile Petunia

Summary Development of anthers in cytoplasmic male sterile (CMS) Petunia diverges from the normal sequence of events early in meiosis. Quantitative and qualitative changes in morphology, proteins and free amino acid contents correlate with this divergence. In anthers of the fertile line (5719), total protein content increases, and SDS-PAGE protein patterns change as the anthers mature. Enhanced levels of three polypeptides with molecular weights of 64,000, 63,000 and 45,000 daltons characterize premeiosis in fertile anthers. Protein levels and patterns from anthers of the CMS line (5707) show little alteration during anther development. Protein synthesis seems to be at least partially blocked in the CMS microspore. The 63,000 and 45,000 dalton proteins are not present, and the absence of any unique protein(s) in the CMS line argues against a virus as the causal agent of CMS in Petunia. Analysis of free amino acids from anthers of the fertile line shows levels of proline and pipecolic acid 2–3 and 10–20 fold higher, respectively, than in the CMS line. The amino acids incorporated into proteins show no such differences; analysis of protein hydrolysates shows similar levels of each amino acid in both fertile and CMS lines at every developmental stage examined.     

Antioxidative responses and proline level in leaves and roots of pea plants subjected to nickel stress

The activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione S-transferase (GST) as well as proline content were studied in leaves and roots of 14 day-old pea plants treated with NiSO₄ (10, 100, 200 μm) for 1, 3, 6 and 9 days. Exposure of pea plants to nickel (Ni) resulted in the decrease in CuZnSOD as well as total SOD activities in both leaves and roots. The activity of APX in leaves of plants treated with 100 and 200 μm Ni increased following the 3rd day after metal application, while in roots at the end of the experiment the activity of this enzyme was significantly reduced. In both organs CAT activity generally did not change in response to Ni treatment. The activity of GST in plants exposed to high concentrations of Ni increased, more markedly in roots. In both leaves and roots after Ni application accumulation of free proline was observed, but in the case of leaves concentration of this amino acid increased earlier and to a greater extent than in roots. The results indicate that stimulation of GST activity and accumulation of proline in the tissues rather than antioxidative enzymes are involved in response of pea plants to Ni stress.