转TaDREB3基因大豆的品质等同性分析

将抗逆基因TaDREB3转入大豆品种东农50,得到3个稳定遗传的株系,并将T4株系种子播于大田中。对3个转TaDREB3基因株系及非转基因对照株系的种子分别进行蛋白质、油份、脂肪酸、异黄酮等品质性状的等同性分析,研究转化外源基因TaDREB3后大豆原有品质性状是否有改变。结果表明：3个转基因株系和对照植株相比,蛋白质含量、油分含量和脂肪酸含量均差异不显著;转基因株系1的大豆黄素虽然和对照差异显著,但是异黄酮总含量和对照没有显著性差异。说明TaDREB3基因的导入并没有改变大豆的品质性状。     

大豆rbcL基因克隆、序列分析及原核表达

利用叶绿体基因组保守性的特征,根据菜豆、豌豆、烟草的rbcL基因序列设计引物,从大豆叶绿体DNA中克隆rbcL基因,全长序列为1488bp,包括1449bp的开放阅读框,编码482个氨基酸。相似性比较显示,此序列与其它10个物种rbcL基因核苷酸的同源性为85．37％～95．31％,氨基酸的同源性为90．87％-96．47％。将该基因与表达载体pET-30a（＋）连接,转化大肠杆菌Rosseta感受态细胞,PCR和酶切鉴定筛选阳性克隆,阳性菌液IPTG诱导后经10％SDS—PAGE分析,结果显示,诱导表达出分子量约为60kD的特异融合蛋白。     

人工microRNAs干扰At3A06基因增强了拟南芥对菌核病的敏感性

基于本研究室自主开发的油菜防御cDNA芯片分析结果,油菜基因Bn3A06受菌核病诱导后特异上调表达。序列比对发现,该基因与拟南芥基因At3A06高度同源,后者编码289个氨基酸,功能未知。本研究用花序浸染法将At3A06基因的人工微RNA（amiRNA）干扰载体（该载体包含一个除草剂草胺磷抗性基因）转化拟南芥。经草胺磷筛选获得3株T1代转基因植株,PCR检测均为阳性;荧光定量PCR检测表明,与野生型对照相比,3个转基因植株的At3A06基因的表达量均显著降低;对其T2代进行菌核病抗性鉴定,结果显示转基因株系对菌核病的敏感性均显著增强（P〈0.05）。推测At3A06基因可能参与了植物的防御反应,与植株对核盘菌抗性相关。     

大豆24kDa油体蛋白与人bFGF融合基因转化拟南芥的研究

为探索大豆油体蛋白与人bFGF（碱性成纤维细胞生长因子）蛋白在植物油体中融合表达的可行性,以大豆总DNA为模板,通过PCR扩增得到大豆油体蛋白基因（Ddoil）及启动子（Ddprm）,构建Ddprm启动的Ddoil基因与植物偏好密码子改造的bFGF基因融合表达载体p1390Ddprm-Ddoil-bFGF,花侵染法转化拟南芥,对T2拟南芥抗性植株基因组进行PCR检测,对种子总蛋白进行SDS-PAGE分析、Western blot检测、ELISA检测及细胞活性测定,为bFGF在其他油料作物的油体系统的表达提供参考。     

Growth and persistence of Listeria monocytogenes isolates on the plant model Arabidopsis thaliana

While the majority of human listeriosis cases appear to be linked to consumption of processed ready-to-eat foods (e.g., deli meats), a few listeriosis outbreaks have been linked to consumption of contaminated vegetables. In this study, we assessed four isolates representing the major Listeria monocytogenes lineages for their abilities to attach to and grow on Arabidopsis thaliana, a well-characterized plant model. When plants were dipped for 5min into 3ml of water containing 8.8logCFU of L. monocytogenes and rinsed repeatedly, L. monocytogenes was recovered from the leaves at densities from 1.52 to 2.17logCFU/cm(2). Ten days after exposure, bacterial numbers had increased over initial numbers by 2.60-2.95logCFU/cm(2). Using L. monocytogenes expressing GFP, bacteria were visualized in the intercellular spaces of A. thaliana leaves, suggesting internalization through stomata. These data indicate that L. monocytogenes can rapidly attach to and multiply on plant surfaces and colonize intercellular spaces in A. thaliana leaves where it may be protected from sanitation treatments. When A. thaliana seeds were exposed to L. monocytogenes, between 4.23 and 4.57logCFU/cm(2) were recovered from leaves 7 days post-germination, suggesting that contaminated seeds can produce contaminated plants. Overall, our study demonstrates that prevention of L. monocytogenes contamination of plants throughout growing stages is critical, consistent with recommendations for other produce-transmitted foodborne pathogens.     

栽培大豆和滩涂野大豆及其杂交后代耐盐性、农艺性状与籽粒品质分析

以栽培大豆和滩涂野大豆杂交组合（N23674×BB52）亲本及其经逐代耐盐性筛选获得的4个F4∶5家系为研究对象,对其苗期耐盐性、农艺性状、籽粒品质进行了分析和比较。结果表明：F44个株系的农艺性状都介于两亲本之间。盐胁迫下F5株系幼苗的耐盐系数、干物质积累和相对生长速率高于母本栽培大豆N23674,其中4076株系最为突出。籽粒品质较其父本BB52种群有明显改善,主要表现在籽粒中蛋白质和多数必需氨基酸、含硫氨基酸含量及氨基酸总量高于双亲,粗脂肪含量介于双亲之间,且接近于其母本N23674;4076和4111株系亚油酸含量超过双亲。     

大豆天冬氨酸途径关键酶基因GmASADH的克隆与分析

本研究基于电子克隆获得的大豆GmASADH基因eDNA序列,根据其编码区设计特异引物,以大豆品种小鹦哥豆总RNA为模板,通过RT—PCR获得了约1130bp的eDNA片段,T／A克隆后进行序列测定。测序结果显示,GmASADH基因家族有两个成员,命名为GmASADHI（FJ581425）和GmASADI-12（HM015631）。GmASADHl编码区长度为1134bp,编码378个氨基酸,由7个外显子和6个内含子构成,定位于Gm08染色体。GmASADH2编码区长度为1131bp,编码377个氨基酸,由7个外显子和6个内含子构成,定位于Gm05染色体。两个基因在氨基酸水平上的相似性达到了96．02％,在二级及三级结构上均有不同程度的差异。ASADH蛋白含有NADB—Rossmannsuper-family和Semialdhyde—dhCsuperfamily结构域。     

普洱茶晒青样游离氨基酸和色泽差异研究

为探讨游离氨基酸和色值在普洱茶晒青样产地及古树茶和台地茶中的差异,分析了来自云南西双版纳州、临沧市普洱茶晒青样中18 种游离氨基酸组分含量,用色差仪测定茶汤色值（L*、a*、b*）,并进行方差分析、主成分分析。结果表明：晒青样中游离氨基酸组分含量在两地间有明显差异,西双版纳样品中茶氨酸、天冬氨酸、谷氨酸、精氨酸等主要鲜味氨基酸含量高于临沧样品。氨基酸主成分分析对西双版纳和临沧晒青样可以很好区分。台地茶游离氨基酸总量、天冬氨酸、谷氨酸、茶氨酸、组氨酸含量均比古树茶高,精氨酸含量比古树茶低,其中氨基酸总量、谷氨酸、精氨酸、组氨酸差异显著,二者也是在主要鲜味氨基酸上存在差异。两地晒青样汤色存在差异,西双版纳样品L*、b*值较临沧大,a*值较小。古树茶与台地茶汤色差异不明显。     

几种鱼鳔的营养成分分析与评价

分析比较了不同鱼鳔的营养成分和氨基酸的含量变化特点，并对鱼鳔的化学组成与功能的关系进行了探讨。结果表明，鱼鳔是一种高蛋白、低脂肪且含有较丰富矿物质的重要水产食品资源。鱼鳔中的营养成分含量因种类不同而变化，不同鱼鳔的脂肪含量变化最大，蛋白质含量的变化最小。氨基酸分析结果为：甘氨酸、丙氨酸、谷氨酸、脯氨酸、精氨酸和天冬氨酸等六种氨基酸构成了鱼鳔蛋白质的优势氨基酸。除了甘氨酸、谷氨酸、脯氨酸和天冬氨酸四种氨基酸在不同鱼鳔中的含量差异较大外，大多数氨基酸的含量变化不大。     

Formate dehydrogenase gene of Arabidopsis thaliana is induced by formaldehyde and not by formic acid

Variability of expression of formate dehydrogenase (FDH) caused by uptake of C-1 compounds was examined by using Arabidopsis thaliana as a model plant. Effects of uptake of several C-1 compounds were evaluated by Northern blot analysis using cDNA of A. thaliana FDH prepared by cloning on the basis of known sequence. As a result, expression of the FDH gene in A. thaliana was not intensely influenced by formic acid, an inherent substrate for FDH, but strongly induced by its reduced form, formaldehyde.     

Nutritional quality of legume seeds as affected by some physical treatments, Part 1: Protein quality evaluation

The effect of water soaking, boiling, roasting, microwave cooking, autoclaving, fermentation and micronization on the nutritional quality of cowpea, pea and kidney bean was investigated. Pea seeds showed the highest crude fat, crude fiber and the lowest moisture contents while kidney bean had the lowest crude fat and the highest crude protein and ash contents. Glu and Asp were the main amino acids in all legumes while Cys and Met contents were the lowest. The protein efficiency ratio (PER), protein chemical score (CS) and the essential amino acid index (EAAI) of raw seeds ranged from 2.15 to 2.95, 39.07 to 68.30, and 55.49 to 62.84, respectively. Soaking, boiling, microwave cooking and autoclaving increased the total essential amino acids in all samples. Furthermore, raw and treated samples showed higher Lys content than the reference protein. Autoclaving was the most effective in improving protein quality followed by micronization, microwave cooking and fermentation. In addition, in vitro protein digestibility was improved after soaking, boiling, microwave cooking, autoclaving and fermentation but was reduced after roasting and micronization.     

Kinetic and mutation analyses of aspartate kinase from Thermus flavus

To reveal the catalytic mechanism of Thermus aspartate kinase, each of 29 amino acid residues that were highly conserved in the sequenced aspartate kinases, was replaced with alanine or leucine by PCR site-directed mutagenesis. Comparison of the kinetic parameters of these mutants with those of the wild-type aspartate kinase suggested that Thr47 was involved in binding aspartate and that Lys7 and Glu74 were involved in catalysis. Analysis of the effective concentrations of magnesium ion on the activity showed that the mutants with replacements at Ser41, Thr47, Asp154 and Asp182 required higher concentrations of magnesium ion. This suggests that these four residues play important roles in the binding of magnesium ions which are required for enzymatic activity.     

Kinetic and mutation analyses of aspartate kinase from thermus flavus

To reveal the catalytic mechanism of Thermus aspartate kinase, each of 29 amino acid residues that were highly conserved in the sequenced aspartate kinases, was replaced with alanine or leucine by PCR site-directed mutagenesis. Comparison of the kinetic parameters of these mutants with those of the wild-type aspartate kinase suggested that Thr47 was involved in binding aspartate and that Lys7 and Glu74 were involved in catalysis. Analysis of the effective concentrations of magnesium ion on the activity showed that the mutants with replacements at Ser41, Thr47, Asp154 and Asp182 required higher concentrations of magnesium ion. This suggests that these four residues play important roles in the binding of magnesium ions which are required for enzymatic activity.     

高γ-氨基丁酸茶游离氨基酸的变化及机理分析

本文对高γ-氨基丁酸（Gabaron）茶加工过程中离体茶鲜叶在无氧条件下游离氨基酸的变化进行了研究，结果表明：常规条件下的离体茶鲜叶游离氨基酸总量为2．5％，其中除含有一般的蛋白质氨基酸外，还含有茶叶特征氨基酸-茶氨酸（The）以及微量的γ-氨基丁酸等。游离氨基酸中含量较高的为谷氨酸（Glu）、脯氨酸（Pro）、天冬氨酸（Asp）、精氨酸（Arg）、组氨酸（His），较低的是苏氨酸（Thr）、丝氨酸（Ser）和甘氨酸（Gly）。离体茶鲜叶经3h无氧处理后，游离氨基酸总量达3．4％。其中Pro、γ-氨基丁酸（Gaba）、Gly、Ser、丙氨酸（Ala）、缬氨酸（Val）、Thr等游离氨基酸含量上升。Pro和Gaba是上升幅度最大的两种氨基酸，分别为正常条件下的3倍和15倍。无氧条件下Glu和ASP含量下降，降幅分别为45％、36％。其余游离氨基酸基本稳定不变。     

奇亚籽营养成分分析及其饮料工艺优化

为开发奇亚籽的营养价值,对黑、白两种奇亚籽的营养成分进行测定,并将其应用在饮料中。采用国标的方法测定基本营养成分,利用氨基酸自动分析仪、气相色谱-质谱联用仪(Gas Chromatography-Mass Spectrometer,GC-MS)分别测定氨基酸及脂肪酸含量,通过单因素实验和响应面试验优化奇亚籽柠檬汁饮料工艺配方。结果表明:黑、白奇亚籽粗蛋白质、粗脂肪、膳食纤维含量分别为16.40%、28.32%、34.67%和29.10%、30.48%、26.65%;二者水分和灰分含量均低于10%。共检出16种氨基酸,黑、白奇亚籽中必需氨基酸含量分别为7.37和7.77 g/100 g,占氨基酸总量的32.92%、38.39%。以氨基酸评分、化学评分为标准,第一限制性氨基酸均为蛋氨酸+半胱氨酸。黑、白奇亚籽中多不饱和脂肪酸分别占脂肪酸总量的82.82%和84.89%,其中omega-3占63.18%~68.37%,具有较高的食用价值。饮料最佳工艺配方:浓缩柠檬汁添加量10%、麦芽糖醇添加量14%、结冷胶添加量0.15%、奇亚籽添加量3%和甜蜜素添加量0.02%。在此条件下,饮料感官评分为(88.5±0.71)分,可制得口感好、色泽明亮、体系稳定均一的奇亚籽柠檬汁饮料。综合分析奇亚籽作为一种新食品原料,其营养价值丰富,并通过优化饮料制作工艺,为其在饮料深加工领域提供理论依据。     

Mutations that cause threonine sensitivity identify catalytic and regulatory regions of the aspartate kinase of Saccharomyces cerevisiae.

The HOM3 gene of Saccharomyces cerevisiae encodes aspartate kinase, which catalyses the first step in the branched pathway leading to the synthesis of threonine and methionine from aspartate. Regulation of the carbon flow into this pathway takes place mainly by feedback inhibition of this enzyme by threonine. We have isolated and characterized three HOM3 mutants that show growth inhibition by threonine due to a severe, threonine-induced reduction of the carbon flow into the aspartate pathway, leading to methionine limitation. One of the mutants has an aspartate kinase which is 30-fold more strongly inhibited by threonine than the wild-type enzyme. The predicted amino acid substitution in this mutant, A406T, is located in a region associated with the modulation of the enzymatic activity. The other two mutants carry an aspartate kinase with reduced affinity for its substrates, aspartate and ATP. The corresponding amino acid substitutions, K26I and G25D, affect residues located in the vicinity of a highly conserved lysine-phenylalanine-glycine-glycine (KFGG) stretch present in the N-terminal part of the aspartate kinase, to which no function has so far been assigned. We suggest that this region is involved in substrate binding. Mutagenesis of a HOM3 region centred in the KFGG-coding triplets generated alleles that determine threonine sensitivity or auxotrophy for threonine and methionine, but not a phenotype associated with a feedback-resistant aspartate kinase, indicating that this region is not involved in the allosteric response of the enzyme.     

不同培养基的平菇多酚及氨基酸含量分析

该研究将不同粉碎处理的葡萄籽和朝鲜蓟叶添加在培养基中,主要研究了工业副产物对平菇营养物质及活性成分的影响.以总酚含量、总黄酮含量、DPPH、ABTS自由基清除能力及Fe3+还原能力为检测指标对其抗氧化活性进行评估;此外,利用相关性分析、主成分分析和偏最小二乘判别等分析手段对平菇氨基酸组成进行探究.结果 表明,培养基中添...     

湘西猕猴桃籽成分分析及猕猴桃籽油的特性研究

分析了湘西猕猴桃籽的主要成分,同时采用水酶法提取猕猴桃籽油,并对猕猴桃籽油的特性进行了研究。结果表明:湘西猕猴桃籽粗脂肪含量32.19%,粗蛋白质含量16.06%;猕猴桃籽中氨基酸种类不全,缺少2种氨基酸,属于不完全蛋白质;鲜味谷氨酸(Glu)和天门冬氨酸(Asp)含量很高,分别为6.1 mg/g和3.5 mg/g,占氨基酸总量的25.98%,为猕猴桃籽增添了良好的风味;水酶法提取猕猴桃籽油总出油率为25.89%,萃取率达80.45%;猕猴桃籽油中亚麻酸含量高达60.59%,不饱和脂肪酸含量达89.92%,生育酚含量为812.41 mg/kg。     

灵芝菌固态发酵夏秋梅占红茶风味特征分析

目的 探究灵芝菌固态发酵下夏秋梅占红茶的风味特征。方法 采用高效液相色谱法(high performance liquid chromatography,HPLC)、电子舌(electronic tongue,E-tongue)与气相色谱-质谱法(gas chromatography-mass spectrometry, GC-MS)相结合,利用多元统计分析,对夏秋梅占灵芝菌茶的滋味与香气品质特征进行判定。结果 与夏秋梅占红茶相比,梅占灵芝菌茶在滋味与香气特征上具有差异, HPLC检测结果显示儿茶素类、咖啡碱类苦涩味物质及部分氨基酸类物质含量显著下降,电子舌结果表明甜味响应值增加,苦味、鲜味响应值降低。其中梅占灵芝菌茶中脯氨酸、没食子酸、儿茶素没食子酸与苦味特性呈显著负相关,天冬氨酸、丙氨酸与涩味特性呈显著负相关,丙氨酸与鲜味特性呈极显著正相关,没食子儿茶素与鲜味特性呈显著正相关。挥发性成分上,建立正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis, OPLS-DA)模型,选取模型中变量重要性投影(v...     

喷施硒肥对大豆理化指标和品质的影响

通过对大豆叶面喷施不同浓度、不同次数的硒肥，研究了硒肥对大豆生理生化指标及品质的影响，结果表明，叶片中非酶类物质可溶性糖和可溶性蛋白含量增加，丙二醛含量减少，保护酶系中过氧化物酶活性、过氧化氢酶活性和超氧化物歧化酶活性增强，对提高植株抵御不良环境的能力有重要的作用；籽粒中粗蛋白含量、粗脂肪含量、氨基酸总量、必需氨基酸总量和硒含量均有增加，明显改善了大豆品质。试验说明，喷施硒肥配制适当浓度与施肥次数相结合能增强大豆作物抗性，改善大豆作物品质。