薇菜、蕺菜中氨基酸及其他营养素含量的测定

对秦巴山区薇菜 (Osmundajaponicathunb .)、蕺菜 (Houttuyniacordatathunb.)两种山野菜可食部分中氨基酸、灰分、粗蛋白、粗脂肪、粗纤维、总糖等含量进行了测定。结果表明 :所测定的 1 7种氨基酸中 ,薇菜含 1 7种氨基酸 ,蕺菜含 1 6种氨基酸 ,七种必需氨基酸含量占各自氨基酸总量的百分数分别为 4 3 .2 6%、3 6.58%。薇菜、蕺菜灰分含量分别为 8.1 1 %、9.4 0 % ;粗蛋白含量分别为 2 2 .80 %、1 7.2 2 % ;粗脂肪含量分别为 3 .3 4 %、5.4 3 % ;粗纤维含量分别为 1 3 .0 7%、1 7.89% ;总糖含量分别为 2 1 .1 4 %、1 3 .4 2 %。     

冀鲁豫花生育成品种的遗传多样性分析

以冀鲁豫三省不同地域的41个花生育成品种为材料,利用SSR分子标记结合田间表型鉴定、聚类分析等方法对其遗传多样性进行了研究。结果表明,21对SSR引物对41个花生育成品种进行了扫描,共检测到52个等位变异,每个位点2～4个,平均2.5个,Shannon信息指数变幅为0.21～1.40,平均为0.73,聚类分析显示在阈值为5.54可将供试品种分为3大类群7个亚类。不同品种9个农艺性状变异系数在13.16%～186.49%之间,基于农艺性状的聚类分析结果显示在阈值为5.48时可以将供试品种分为6大类群11个亚类,各大类群间品种的农艺性状表现各具特点。     

人工发酵古尼虫草中甘露醇的测定

利用比色法简便、准确和快速地测定发酵生产的古尼虫草菌丝中甘露醇含量。结果表明 ,甘露醇的最大特征吸收峰在 41 2nm处 ,质量浓度在 1 0～ 5 0mg/L范围内线性较好 ,其回归方程 ρ=-0 842 +97 3 2 9A,r=0 9992 ;平均回收率 1 0 0 2 4% ,RSD =0 5 4% (n=5 )。样品中甘露醇采用水提取法 ,提取时间 2h。甘露醇显色后 2 0min内测定对结果影响不大 ,在测定时要注意排除果糖对测定结果的影响。用此法测得发酵古尼虫草菌丝体中甘露醇的含量为 7.4% ,RSD =0 .2 4% (n =6)。     

薏苡种子形态性状多样性评价

薏苡是优质的药食兼用作物。本研究对我国36份薏苡种质的种子形态性状进行了主成分分析和聚类分析。结果表明,36份薏苡种质具有较高的形态性状多样性。不同薏苡种质的4个种子质量性状间存在差异。单因素方差分析显示11个种子数量性状在不同薏苡种质间呈极显著差异。数量性状的变异系数在8.43%~68.01%之间,种壳与外种皮重量的变异系数最大而种仁长度的变异系数最小。主成分分析将36份薏苡种质分为中部及南部栽培种质、北部栽培种质和野生种质3大类,3类种质的形态特征存在明显差异。聚类分析将36份薏苡种质分为5大类群,即中部及南部栽培种质、北部栽培种质和3个类群野生种质。初步筛选出6份可能具有较高抗肿瘤活性的薏苡种质FJNJ、TJ-BK-1、TJ-BK-2、MY-BK-1、MY-BK-2和JYSD。     

嗜酸乳杆菌P302菌株产类细菌素最佳条件的研究

目的对产类细菌素嗜酸乳杆菌P302的发酵条件进行研究。方法采用琼脂扩散法测定发酵上清液对大肠埃希菌O139的抑菌活性。结果通过正交试验确定P302产类细菌素的最佳培养基组分为牛肉膏4％、胰蛋白胨0．2％、酵母粉0．8％、碳酸钙0．4％和蔗糖1％,0．4％复合维生素,0．3％Tween-80有利于类细菌素的产生。发酵工艺条件为最适初始pH7．0,最适温度37℃,最佳接种量0．3％,种龄14h,厌氧培养22h。结论通过优化发酵条件和发酵工艺,类细菌素的产量提高了35％。     

红麻品种(系)表型性状的因子和聚类分析

依据红麻高产育种目标,利用因子分析和聚类分析对40个红麻品种(系)的9个表型性状进行分析。结果表明:供试材料9个表型性状的变异系数为5.13%~25.33%,具有较大差异和丰富的遗传多样性;对供试红麻材料进行聚类分析,可分为3大类,其中,第Ⅲ类群属理想的高产品系。二维排序分析,评选出7个综合性状表现较好的亲本,T17-2、福红951、FH航992、H1301、ZHKX-01、福航优1号、福航优3号共7个品系,可在生产和育种中加以利用。     

水稻和拟南芥中几丁质酶的分析

几丁质酶（EC3．2．1．14）是一种降解几丁质的糖苷酶,广泛存在于各种生物体中,并在植物中对病原真菌起重要抗性作用。首先通过BLAST在GenBank中对其同源性进行搜索,用SMART分析其结构。基于水稻和拟南芥的基因组注释,借助4个生物学软件（SignalP3．0,TMHMM2．0,TargetP1．1andbig—PiPredictor）,分析了水稻所有37条和拟南芥所有24条几丁质酶序列,发现有些几丁质酶都分泌到细胞外,有些定位于液泡中,水稻中仅25条和拟南芥中仅16条几丁质酶序列有信号肽,这些信号肽的平均长度为24．8aa。利用ClustalX和MEGA3．1两个生物软件分析了59条几丁质酶序列和25条标准几丁质酶的系统发育关系,这些几丁质酶可分为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ和Ⅵ等6大类。这种聚类结果与几丁质酶传统分为7类有些差异。通过对6大类中各个氨基酸残基的分析,发现丙氨酸、甘氨酸、丝氨酸和亮氨酸的使用频率在每类中都非常高,而蛋氨酸、组氨酸、色氨酸和半胱氨酸均低于20％。各大类中彼此之间的某些氨基酸使用频率明显不同,Ⅰ-Ⅵ分别富含丙氨酸、缬氨酸、亮氨酸、半胱氨酸、丝氨酸和赖氨酸。     

小麦抗赤霉病品种NBS同源序列克隆与分析

根据二穗短柄草NBS-LRR类基因的保守序列设计同源引物,以小麦抗赤霉病品种苏麦3号、宁7840和望水白基因组DNA为模板,通过PCR扩增,得到43条序列,其中4条为非编码序列或结构域不完整;39条与植物抗病基因同源,其中的7条内部存在终止密码子,可能是假基因,经过比对分析,其余32条具有连续的开放阅读框和保守结构域,推导的氨基酸序列均具有Kinase-1a、Kinase-2和Kinase-3a及GLPL区等几个保守区,在GenBank中均能找到与之高度同源的其他物种的核酸序列,并且Kinase-2的最后一个氨基酸均为色氨酸(W),属于non-TIR类NBS基因。32条序列可分为4大类,它们之间核苷酸同源性为64%-98%,编码氨基酸同源性为22%-98%。根据序列分析随机设计5对不同基因特异性引物,并利用RT-PCR技术进行表达分析,结果表明,7-1、s-3、s-4和w-2均能表达,说明这些片段可能是功能性抗病基因的部分序列;7-13不表达,再次证明属于假基因。32条序列在之前未被报道过,这些RGA可以作为筛选赤霉病功能性抗病基因的候选序列。     

山荆子腊叶标本表型性状变异分析

为了揭示山荆子表型性状的变异程度和规律,以150份山荆子腊叶标本为材料,选择20个表型性状,统计基本参数并进行聚类分析.结果表明:叶的平均变异系数(CV=32.67%)大于果的平均变异系数(CV=20.7%),果的性状稳定性较高,叶性状中叶柄长变异系数(CV=38%)最大,叶宽位/叶长变异系数(CV=13.29%)最小;果实性状中果径宽位/果柄长(CV=29.37%)变异最大,变异最小的性状是果长/果径(CV=13.10%);选用平均欧氏距离对各地方居群的20个表型性状进行UPGMA聚类,在距离5阈值处,可以划分为5类,在欧氏距离6阈值处,可划分为3大类.     

基于表型参数及SRAP标记的广东茶树种质遗传多样性

采用表型鉴定与SRAP分子标记,对25份广东茶树种质和5份对照品种的遗传多样性进行系统评价和分类;利用Pearson相关和Farthest neighbor方法对其表型性状进行聚类分析.结果表明:茶树各性状的平均变异系数为32.15%,其中茸毛的变异系数最大,为42.41%;芽叶生育期变异系数最小,为18.52%.经基于表型性状的聚类分析,30份样本可分为4组：第1组有17个品种,第2组有10个品种,第3组为云南大叶种和凌云白毛茶2个对照品种,第4组仅海南大叶种1个对照品种.利用21对SRAP引物对茶树基因组DNA进行研究,共扩增出127条带,其中114条为多态性带,占88.67%;平均每个引物组合的谱带数和多态性带数分别为6.05条和5.43条.当遗传距离为0.39 cm时,茶树种质可分成A、B、C 3类,其中A类占83.33%;当遗传距离为0.31 cm时,又可将A群划分为Ⅰ、Ⅱ、Ⅲ3个亚群,其中第Ⅰ亚群包括13个品种,第Ⅱ亚群包括2个品种,第Ⅲ亚群包括10个品种.依据SRAP标记的聚类与表型性状的表现并不完全一致.     

More monensin-sensitive, ammonia-producing bacteria from the rumen.

Two monensin-sensitive bacteria which utilized carbohydrates poorly and grew rapidly on amino acids were isolated from the bovine rumen. The short rods (strain SR) fermented arginine, serine, lysine, glutamine, and threonine rapidly (greater than 158 nmol/mg of protein per h) and grew faster on casein digest containing short peptides than on free amino acids ().34 versus 0.29 h(-1)). Gelatin hydrolysate, an amino acid source containing an abundance of long peptides, was unable to support growth or ammonia production, but there was a large increase in ammonia production if strain SR was cocultured with peptidase-producing ruminal bacteria (Bacteroides ruminicola or Streptococcus bovis). Cocultures showed no synergism with short peptides. Strain SR washed out of continuous culture ().1 h(-1)) at pH 5.9. The irregularly shaped organisms (strain F) deaminated glutamine, histidine, glutamate, and serine rapidly (greater than 137 nmol/mg of protein per min) and grew faster on free amino acids than on short peptides ().43 versus 0.21 h(-1)). When strain F was provided with casein or gelatin hydrolysate and cocultured with peptidase-producing bacteria, there was a more than additive increase in ammonia production. Strain F grew in continuous culture (0.1 h(-1)) when the pH was as low as 5.3. The irregularly shaped cells and short rods were present at less than 10(9)/ml in vivo, but they ahd very high specific activities of ammonia production (greater than 310 nmol of ammonia/mg of protein per min) and could play an important role in ruminal amino acid fermentation.     

More monensin-sensitive, ammonia-producing bacteria from the rumen

Two monensin-sensitive bacteria which utilized carbohydrates poorly and grew rapidly on amino acids were isolated from the bovine rumen. The short rods (strain SR) fermented arginine, serine, lysine, glutamine, and threonine rapidly (greater than 158 nmol/mg of protein per h) and grew faster on casein digest containing short peptides than on free amino acids ().34 versus 0.29 h(-1)). Gelatin hydrolysate, an amino acid source containing an abundance of long peptides, was unable to support growth or ammonia production, but there was a large increase in ammonia production if strain SR was cocultured with peptidase-producing ruminal bacteria (Bacteroides ruminicola or Streptococcus bovis). Cocultures showed no synergism with short peptides. Strain SR washed out of continuous culture ().1 h(-1)) at pH 5.9. The irregularly shaped organisms (strain F) deaminated glutamine, histidine, glutamate, and serine rapidly (greater than 137 nmol/mg of protein per min) and grew faster on free amino acids than on short peptides ().43 versus 0.21 h(-1)). When strain F was provided with casein or gelatin hydrolysate and cocultured with peptidase-producing bacteria, there was a more than additive increase in ammonia production. Strain F grew in continuous culture (0.1 h(-1)) when the pH was as low as 5.3. The irregularly shaped cells and short rods were present at less than 10(9)/ml in vivo, but they ahd very high specific activities of ammonia production (greater than 310 nmol of ammonia/mg of protein per min) and could play an important role in ruminal amino acid fermentation.     

The Nutrition of Parauronema acutum*

SYNOPSIS. A symbiote-free strain of Parauronema acutum, 110–3, a small marine hymenostome ciliate has been cultured in a synthetic medium consisting of amino acids, purine derivatives, vitamins, lipids and artificial sea water. Populations of ～ 1.3 × 10⁶ per ml were obtained in 5 to 6 days at 27 C in the dark in medium prepared in sea water, density = 1.015 g/cc at a surface to volume ratio of 5 cm²/ml. The pH optimum was 7.2. The following amino acids were determined to be essential for the growth of this strain: arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine (or glycine), threonine, tryptophan and valine: guanine, guanosine or guanylic acid, but not adenine, adenosine, adenylic acid, hypoxanthine, inosine, inosinic acid, xanthine, xanthosine, or xanthylic acid, satisfied the need for a purine for growth of this organism. Pyrimidines were not required for growth. Of the vitamins tested, folic acid, nicotinamide, d-pantothenic acid, pyridoxal HCl, riboflavin, thiamine HCl and thioctic acid were essential for growth; biotin was not. Growth in the absence of lipids was transplantable, but amounted to ～ 3% that obtained in medium containing a mixture of asolectin, animal cephalin and Tween 80. Asolectin alone at high concentrations was almost as effective as the lipid mixture in supporting growth. Purified phospholipids such as phosphatidyl serine, phosphatidyl choline and phosphatidyl inositol were less effective on an individual basis. In minimal medium containing only the “essential” amino acids, growth was less than 5% that obtained in the complete medium, but could be restored to maximal by the addition of either glutamic acid or aspartic acid. A number of substances, including sugars, amino acids and Krebs cycle intermediates, partially restored growth under these conditions. Only glycogen, starch and glucose-1-phosphate, tested individually, were as effective as glutamic acid or aspartic acid in restoring growth to optimal levels.     

Complete amino acid analysis of peptides and proteins after hydrolysis by a mixture of Sepharose-bound peptidases

Incubation with a mixture of Sepharose-bound peptidases was shown to result in the quantitative release of amino acids from certain peptides and S-aminoethylated proteins. Subtraction of the low background values of amino acids generated by the enzymes enables amino acid ratios of corticotrophin-(1-24)-tetracosapeptide to be determined with a standard deviation on repeat digestions of 3-5%. Good values were obtained for amino acids that are completely or partially destroyed on acid hydrolysis, i.e. tryptophan, tyrosine, serine, asparagine and glutamine. Experiments with peptides containing d-amino acids showed that the enzyme mixture is stereospecific and could therefore be used to detect the presence of d-residues in peptides. The enzyme mixture completely hydrolyses peptide fragments obtained after Edman degradation and should therefore be useful for determining sequences of peptides containing acid-labile amino acid residues. The activities of the bound enzymes were unaltered over a period of 7 months and they provide a simple, reproducible procedure for the quantitative determination of amino acids in peptides and proteins containing l-amino acids.     

Regulation of RNA degradation in cultured rat hepatocytes: Effects of specific amino acids and insulin

The regulation of RNA degradation by specific amino acids and insulin was investigated in cultured rat hepatocytes from fed rats previously injected in vivo with [6-¹⁴C]orotic acid. The effects of three groups of amino acids were compared to those of a complete amino acid mixture. The first one consisted of the eight amino acids (leucine, proline, glutamine, histidine, phenylalanine, tyrosine, methionine, tryptophan) previously found to be particularly effective in the control of proteolysis. The two other groups were defined from our study with single additions of amino acids, one consisting of proline, asparagine, glutamine, alanine, phenylalanine, and leucine and the other including the latter group with serine, histidine, and tyrosine. The results showed that these three groups were able to strongly inhibit deprivation-induced RNA breakdown at one and ten times normal plasma concentrations but to a lower extent than the complete amino acid mixture. Six amino acids (proline, asparagine, glutamine, alanine, phenylalanine, leucine) inhibited individually RNA degradation by more than 20%. However, the deletions of proline, asparagine, glutamine, or alanine from the group of these six amino acids were not followed by a loss of inhibitory effect. On the contrary, an important loss of inhibition was observed when leucine and phenylalanine were deleted. Furthermore, only these two amino acids exhibited an additive inhibitory effect. Thus leucine and phenylalanine could be considered as important inhibitors of RNA breakdown in cultured rat hepatocytes. Finally, insulin which had no significant effect on RNA degradation in the absence of amino acids, was able to potentiate the inhibitory effect of different amino acid groups. © 1993 Wiley-Liss, Inc.     

The phosphorylated pathway of serine biosynthesis links plant growth with nitrogen metabolism

Because it is the precursor for various essential cellular components, the amino acid serine is indispensable for every living organism. In plants, serine is synthesized by two major pathways: photorespiration and the phosphorylated pathway of serine biosynthesis (PPSB). However, the importance of these pathways in providing serine for plant development is not fully understood. In this study, we examine the relative contributions of photorespiration and PPSB to providing serine for growth and metabolism in the C3 model plant Arabidopsis thaliana. Our analyses of cell proliferation and elongation reveal that PPSB-derived serine is indispensable for plant growth and its loss cannot be compensated by photorespiratory serine biosynthesis. Using isotope labeling, we show that PPSB-deficiency impairs the synthesis of proteins and purine nucleotides in plants. Furthermore, deficiency in PPSB-mediated serine biosynthesis leads to a strong accumulation of metabolites related to nitrogen metabolism. This result corroborates ¹⁵N-isotope labeling in which we observed an increased enrichment in labeled amino acids in PPSB-deficient plants. Expression studies indicate that elevated ammonium uptake and higher glutamine synthetase/glutamine oxoglutarate aminotransferase (GS/GOGAT) activity causes this phenotype. Metabolic analyses further show that elevated nitrogen assimilation and reduced amino acid turnover into proteins and nucleotides are the most likely driving forces for changes in respiratory metabolism and amino acid catabolism in PPSB-deficient plants. Accordingly, we conclude that even though photorespiration generates high amounts of serine in plants, PPSB-derived serine is more important for plant growth and its deficiency triggers the induction of nitrogen assimilation, most likely as an amino acid starvation response.

The phosphorylated pathway of serine biosynthesis is required to synthesize serine for plant growth; and its deficiency triggers an amino acid starvation response by inducing nitrogen assimilation.     

Quantitative analysis of free amino acids and carbohydrates in spring barley anthers at different stages of maturity

The content of the carbohydrates glucose, fructose and sucrose was determined in spring barley anthers at different stages of maturity. During maturation the sucrose content of the anthers increased markedly. The following 17 free amino acids were detected in anthers of different stages of maturity: aspartic acid, glutamic acid, serine, alanine, arginine, leucine, isoleucine, lysine, α-aminobutyric acid, glutamine, proline, tyrosine, phenylalanine, valine, threonine, cystine and glycine. Quantitative analysis was only carried out in amino acids present in higher concentrations in the analysed samples. These were: aspartic acid, glutamic acid, α-aminobutyric acid, proline, serine, valine and glutamine, and a mixture of amino acids (leucine, isoleucine, valine and phenylalanine). The total content of free amino acids increased with increasing maturity of the anthers. However, not all amino acids followed contributed to this increase, but only proline, glutamic acid, aspartic acid and glutamine. A small difference was found in the variety Gopal in which the aspartic acid content did not increase significantly, but the content of the mixture of amino acids and serine did. With the exception of green anthers of the variety Firlbecks Union, proline was present in the highest concentration in all samples analysed.     

Regulation of glutamine synthesis by glycine and serine in Neurospora crassa.

The biosynthetic activities of the polypeptide subunits alpha and beta of glutamine synthetase (GS) were inhibited in vitro by glycine and serine. These amino acids inhibited the growth of a mutant strain with partial GS activity when grown on glutamate as the nitrogen source and also blocked the synthesis of the glutamine in vivo, thus demonstrating the inhibitory effect on GS activity in vivo. Glycine and serine lowered the intracellular glutamine pool and regulated GS beta synthesis. A preferential induction of synthesis of the GS beta polypeptide was observed when either of these amino acids was present in the medium. On this basis, we obtained a glycine-sensitive mutant which showed a structural alteration of the GS beta polypeptide. The double regulatory effect of either glycine or serine on glutamine synthesis may be considered an example of the regulation of glutamine synthesis by alpha-amino nitrogen. It may be a mechanism that regulates the assimilation of ammonium into glutamate versus glutamine.     

Amino acid utilization by L-M strain mouse cells in a chemically defined medium

Summary The amino acid requirements of strain L-M mouse cells grown in a chemically defined medium (2×Eagle) containing only the 13 essential amino acids (EAA) were investigated. Medium and acid hydrolysate samples were analyzed for amino acid content by the method of ion exchange chromatography. The extent of utilization of the EAA differed;e.g. after 120 hr of cell growth without medium change, glutamine was exhausted from the medium; methionine, leucine, isoleucine, cystine, arginine, and valine were depleted 60 to 80%; other EAA were used to lesser extents. Although the EAA were used in excess of their requirements for protein synthesis, a correlation could generally be made between utilization and protein amino acid composition. Glutamine appeared to be, a growth-limiting factor. Use of U-¹⁴C-labeled glutamine indicated that over one-half of the metabolized glutamine was converted to carbon dioxide, 17% to cell material, and 15% was extracted from the amino acid pools. Nonessential amino acids (NEAA), viz. alanine, aspartic acid, glutamic acid, glycine, proline, and serine, were released into the medium during growth, and some were reutilized. Exogenous provision of these did not improve cell growth. In contrast to the other NEAA, only serine showed net utilization when provided exogenously. When glutamic acid largely replaced the glutamine in the medium, it exerted a sparing effect on the glutamine requirement for protein synthesis. Suggestions are given for the improvement of Eagle medium for cell growth. Supported by Research Grants CA 03720 and CA 11802 from the National Institutes of Health. Predoctoral, fellow supported, by Grant F01-GM-42156-02 from the National Institutes of Health.