红花脂质转运蛋白基因的克隆及表达分析

克隆红花脂质转运蛋白基因(Lipid transfer protein,LTP),并进行生物信息学及表达分析,旨为研究LTP在红花抵抗逆境胁迫中的作用提供依据。通过RT-PCR方法从红花种子中克隆LTP基因序列,通过生物信息学对该基因蛋白的特征进行分析,构建LTP与相关物种LTP的系统进化树,利用Real-time PCR方法分析在红花不同号组织中LTP基因的表达量。结果显示,LTP基因ORF全长294 bp,编码97个氨基酸,相对分子量为7.46 kD,等电点为8.91。红花LTP蛋白包含一个长为29个氨基酸残基的信号肽序列;该蛋白含有一个丝氨酸磷酸化位点;三级结构预测表明该蛋白是由3个α-螺旋和一个β-转角简单的缠绕在无规则卷曲上的简单结构。分子进化表明,红花LTP基因与十字花科的甘蓝型油菜进化关系最近。通过荧光定量PCR对红花LTP基因的组织表达特异性进行分析,结果表明Ct LTP在不同组织的表达水平具有显著差异,在种子和花中呈现高表达,而在其他组织中低表达。     

红花脂质转运蛋白基因的克隆及表达分析北大核心CSCD

克隆红花脂质转运蛋白基因(Lipid transfer protein,LTP),并进行生物信息学及表达分析,旨为研究LTP在红花抵抗逆境胁迫中的作用提供依据。通过RT-PCR方法从红花种子中克隆LTP基因序列,通过生物信息学对该基因蛋白的特征进行分析,构建LTP与相关物种LTP的系统进化树,利用Real-time PCR方法分析在红花不同号组织中LTP基因的表达量。结果显示,LTP基因ORF全长294 bp,编码97个氨基酸,相对分子量为7.46 kD,等电点为8.91。红花LTP蛋白包含一个长为29个氨基酸残基的信号肽序列;该蛋白含有一个丝氨酸磷酸化位点;三级结构预测表明该蛋白是由3个α-螺旋和一个β-转角简单的缠绕在无规则卷曲上的简单结构。分子进化表明,红花LTP基因与十字花科的甘蓝型油菜进化关系最近。通过荧光定量PCR对红花LTP基因的组织表达特异性进行分析,结果表明Ct LTP在不同组织的表达水平具有显著差异,在种子和花中呈现高表达,而在其他组织中低表达。     

两性花双优山葡萄果实氨基酸的研究

作者通过对两性花双优山葡萄果实氨基酸的研究,从试验得知：同一品种在不同地区栽培其氨基酸的总量及各种氨基酸量不同。但在各地方果实中氨基酸的分配基本规律是一致的：种子含量为最高,果皮其次,果肉最低。各部位氨基酸的大小值：种子为１０．４４５—８．４７５,果皮为６．４０７—２．８８２,果肉为４．６１８—２．５６４。     

东北地区三种柴胡种子游离氨基酸的研究

东北地区三种柴胡１２个居群做了种子游离氨基酸成分分析。发现狭叶柴胡９个居群中种子游离氨基酸种内存在着变异,而且种间游离氨基酸种类变异较大,可以作为种间分类的指标。     

磁化水对红花种子萌发的影响

磁化水,亦名磁水、磁处理水,是指水经磁场作用,水的理化性能发生变化,保持有生物效应的活性水。关于磁化水对某些植物种子萌发及幼苗生长的影响,国内外已有一些报导,但由于磁场场型、场强及实验条件等不同,其结果也不一样。对中药红花种子的影响,国内报道甚少。本实验根据磁化水产生生物磁效应之原理,选用0.2T场强磁水器对红花种子进行处理,旨在探讨该场强磁化水,对红花种子萌发的影响程度。结果表明:红花种子经磁化水处理后发芽指标明显提高。材料与方法试验材料:红花种子,购于内蒙古蒙药厂。用SH型磁水器,上饶磁性材料厂制,场强0.2T。磁…     

大豆油体与EGF融合基因的克隆及其在红花种子中的表达

目的：旨在建立一种在红花油体中表达EGF的转基因植物的方法。方法：通过PCR技术把大豆油体基因（DDoil）与EGF构建成融合基因,克隆至植物表达载体pCAMBIA1390R中,构建成植物表达载体p1390Do-EGF,然后转化进农杆菌 LBA4404 中用于侵染红花外植体,通过甘露糖筛选培养基培养可获得红花转化苗。通过PCR、实时荧光相对定量RT-PCR、SDS-PAGE和Western blot分析目的基因的表达情况,通过MTT法检测EGF的促细胞增殖活性。结果：PCR结果显示,红花叶片中能检测到EGF基因;实时荧光相对定量RT-PCR结果显示,在红花种子中EGF能成功实现转录;SDS-PAGE和Western blot检测证明,在转基因红花种子中能有效表达出EGF,并具有其原有的免疫原性,MTT法实验结果表明EGF具有促进balb/c 3T3细胞增殖的生物活性。结论：大豆油体和EGF融合基因已经成功转化进红花细胞的基因组中,并实现了EGF外源蛋白在红花种子油体中的表达,为EGF蛋白的产业化生产探讨了一种新的生产途径。     

超干红花种子抗老化作用及其机理

红花种子含水量（ＭＣ）降至５％以下,能显著提高抗老化劣变能力;在同等老化条件下,对照种子（ＭＣ＝９．０１％）活力已大幅度下降,而超干种子的活力保持不变,超干处理对红花种子脱氢酶,过氧化氢酶（ＣＡＴ）及超氧物歧化酶（ＳＯＤ）活性均无明显影响。经同等老化处理后,超干种子脱氢酶活性明显高于对照种子。在同等老化条件下,当对照种子ＣＡＴ和ＳＯＤ活性明显下降时,超干种子仍保持较高活性。红花种子在超干老化处理后     

六种彩叶植物营养成分及有害元素含量分析

以六种彩叶植物紫叶李、红花木、红叶石楠、红枫、鸡爪槭和金叶女贞的叶片为材料,对其营养成分及有害元素含量进行了测定及分析。结果表明:蛋白质和可溶性糖含量以红花木为最高,维生素C含量以红枫为最高,β-胡萝卜素含量差异最大,紫叶李最高达15.99mg·kg-1。六种彩叶植物矿质营养元素种类齐全,其中红花木的营养元素总量最高。六种彩叶植物均含有18种氨基酸成分,必需氨基酸和总氨基酸含量各不相同,但必需氨基酸占总氨基酸的比例却较为一致,且都高于40%。六种彩叶植物叶片内均含有一定量的铅、镉、砷、汞等有害元素,但有害元素尤其是砷和汞的含量均低于允许量。     

贵州五种野山茶种子氨基酸及脂肪酸成分含量的研究

采用高效液相色谱(HPLC)和氨基酸比值系数法,对贵州五种野山茶种子氨基酸和主要脂肪酸成分含量进行了研究。结果表明:(1)五种野山茶种子中均含有13种氨基酸,不同种类的氨基酸总量差异极显著,必需氨基酸和非必需氨基酸含量的差异显著(P0.01,P0.05),且第一限制性氨基酸含量不同;长柱红山茶(Camellia longistyla)和贵州红山茶(C.kweichouensis)的氨基酸总量及人体必需氨基酸含量分别为33.01、13.29和26.33、10.38 mg·g~(-1)。(2)不同种类种子的千粒重、含油率、含水率皆呈显著差异(P0.01,P0.05),种子含油率与不饱和脂肪酸含量显著正相关(R=0.556、P0.05),种子不饱和脂肪酸含量与棕榈酸、饱和脂肪酸含量极显著负相关(R=-0.791,P0.01;R=-0.776,P0.01),其中长柱红山茶和贵州红山茶种子的含油率和不饱和脂肪酸含量分别为43.93%、71.89%和43.91%、71.85%。(3)小黄花茶(C.luteoflora)、皱叶瘤果茶(C.rhytidophylla)、贵州红山茶、长柱红山茶、美丽红山茶(C.delicata)所含必需氨基酸与总氨基酸比值分别为44.9、43.3、39.0、40.9、33.8,必需氨基酸与非必需氨基酸比值分别为81.6、76.4、64.0、67.4、51.0,除了美丽红山茶的比值系数偏低外,其余4种均达到了WHO/FAO的理想蛋白质标准,种子蛋白质均具有较高的营养价值,其中长柱红山茶和贵州红山茶的种子含油脂质量等级指标接近我国现行茶油标准(GB11765—2003),可以作为重要的优质油茶种质资源加以开发利用。     

果树种子氨基酸含量比较

果树种子中17种氨基酸总量以仁果类最高,其次是核果类、浆果类和坚果类。种子中人体必需的八种氨基酸占总量的比率,浆果高于坚果、核果、仁果。种子含谷氨酸最多,天门冬氨酸、精氨酸、蛋氨酸等次之,组氨酸、丙氨酸、胱氨酸最少。     

Chemical differences between seeds and elaiosomes indicate an adaptation to nutritional needs of ants

Ant-dispersed plants usually produce seeds with appendages (elaiosomes) as reward for ants. Plants that produce high-quality elaiosomes benefit because ants preferentially disperse their diaspores. We therefore hypothesized that seeds and elaiosomes differ in chemical composition in ways that make elaiosomes of high nutritional quality for ants, capable of providing essential dietary components that explain the increased fitness and higher gyne production documented for colonies with elaiosome consumption. To test the hypothesis we analysed the content and composition of lipids, amino acids, soluble carbohydrates, proteins and starch in seeds and elaiosomes of 15 central European ant-dispersed plants. After separating the different fractions, total lipids were determined gravimetrically, fatty acids and soluble carbohydrates were detected by gas chromatography (GC) and GC–mass spectrometry, free amino acids by an amino acid analyser while starch and protein were analysed photometrically. Seeds accumulated high molecular weight compounds such as proteins and starch, whereas elaiosomes accumulated more easily digestible low molecular weight compounds such as amino acids and monosaccharides. Analysis of similarities and similarity percentages analysis demonstrated that the composition of fatty acids, free amino acids and carbohydrates differed markedly between elaiosomes and seeds. The most important difference was in total amino acid content, which was on average 7.5 times higher in elaiosomes than in seeds. The difference was especially marked for the nitrogen-rich amino acid histidine. The availability of essential nutrients and, in some species, the higher nitrogen content in elaiosomes suggest that their nutritional value for larvae plays a key role in this interaction. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The importance of green seed in the nitrogen nutrition of the Zebra Finch Taeniopygia guttata

Abstract Australian grass-finches are widely reported to consume large quantities of green seed when it becomes available, and the opportunistic breeding of wild Zebra Finches in the arid zone has been correlated with the occurrence of rain. In this study, green and ripe seeds were harvested from seven pasture and weed grasses grown in experimental plots and, along with three cereal flours and whole-egg powder, were analysed for the amino-acid composition of their protein. The relative levels of ten amino acids essential in the diets of growing birds were compared between samples using a cluster analysis dendrogram generated from Raabe's Similarity Index. The protein of all green seeds clustered with whole egg, and away from all but one of the ripe seeds and seed products. Green and ripe seed profiles were found to be significantly different by a two-sample multivariate test of significance (Hotelling's T²). Histidine, lysine, phenylalanine and threonine were the amino acids most different. Of these four amino acids, lysine and threonine (along with methionine) were potentially limiting in ripe seeds when compared with whole-egg protein. In green seeds, lysine was only marginally limiting, threonine was no longer limiting, but methionine was still limiting when compared with whole-egg protein. These results indicate that the benefit of green vs ripe seed in the diet of breeding Zebra Finches is partly a higher level of the limiting essential amino acid, lysine, and partly a higher intake and throughput of soft green seed and consequent greater extraction of limiting essential amino acids.     

大麦籽粒及花药愈伤组织的游离氨基酸含量分析

对同一基因型大麦花药愈伤组织与籽粒中的游离氨基酸含量尤其是赖氨酸含量之间的关系进行分析,结果表明,对同一基因型而言,愈伤组织及籽粒中的同一氨基酸含量高低具有相同趋势。同时着重分析了愈伤组织中游离脯氨酸的含量与绿苗分化的关系,结果表明游离脯氨酸含量高的愈伤组织,其绿苗分化率较高,说明脯氨酸对绿苗分化具有重要作用。966259的花药愈伤组织及籽粒中的游离氨基酸总含量及游离赖氨酸含量均最高。     

Pitfalls in using sodium hypochlorite as a seed disinfectant in C incorporation studies

Seeds sterilized with sodium hypochlorite (NaOCl) retained sufficient amounts to interfere with studies of amino acid metabolism of the sterilized seeds during germination. Repeated washing in water did not remove NaOCl completely. However, soaking the seeds for 10 min in 0.01 n HCl removed NaOCl completely, without reducing germinability.Residual NaOCl reacted with the amino acids and reduced their concentrations in the incubation media. This reaction resulted in high production of CO(2) and low uptake of amino acids by the seeds. Decarboxylation of the amino acids occurred in the incubation medium outside the seed, was independent of the presence of seeds in the reaction, and therefore was not related to amino acid metabolism by the seeds. Effects of NaOCl on uptake, incorporation, and CO(2) production from indoleacetic acid were similar to those of the amino acids studied.     

Protein and free amino acids in field-grown cowpea seeds as affected by water stress at various growth stages

Summary This study was undertaken to evaluate water stress effects during vegetative, flowering, and podfilling stages of cowpea plants (Vigna unguiculata L.) grown under natural field conditions in southern California on seed yield and protein and free amino acid content of the cowpea seeds. The lowest concentration of N was found in the seeds of the control treatment plants while the seed yield from these treatments was the highest as compared with the N concentration and yield of seeds from plants subjected to water stress during flowering and podfilling stages. The concentration of N in the seeds was inversely related to the seed dry weight yield. Protein arginine,-threonine,-serine,-cystine,-valine,-methionine, and-isoleucine were significantly affected by water stress at the three growth stages. There was no consistent pattern in the effect of water stress on the individual amino acids. The sum of protein amino acids in the cowpea seeds was not significantly influenced by the various treatments since some of the protein amino acids increased and others decreased producing an averaging effect on the figures comprising the sums of the amino acids. Water stress during the flowering and pod-filling stages increased the free amino acid pool, and at the same time, inhibited incorporation of the amino acids into the protein chain-thus lowering the protein amino acid fraction simultaneously. With the exception of methionine plus cystine, the essential amino acids in the seeds were present at concentrations equal to or greater than recommended by the World Health Organization and FAO. It is of particular importance to note that the concentration of lysine in the cowpeas was substantially higher than that found in wheat grain. It is also important to note that the amount of essential amino acids per gram of protein was not measurably affected by the water stress treatments during any of the growth stages.     

In-depth genome diversity,population structure and linkage disequilibrium analysis of worldwide diverse safflower (Carthamus tinctorius L.) accessions using NGS data generated by DArTseq technology

Molecular Biology Reports - Safflower (Carthamus tinctorius L.) is one of the most important oilseed crops for its seed oil rich in unsaturated fatty acids. Precise utilization of diverse genetic...     

Dormancy in Seeds of Charlock (Sinapis arvensis L.): Early Effects of Gibberellic Acid on the Synthesis of Amino Acids and Proteins

Charlock (Sinapis arvensis L.) seeds were imbibed with 10 mm GA(3) for 24 hours at 0 C. After equilibration at 25 C, a 5-fold increase in radioactivity in the amino acids labeled from 2-(14)C-acetate was observed within 2 hours. The total amount of amino acids was reduced to half, and the specific radioactivity increased approximately 10-fold, indicating a diversion of metabolites for amino acid and protein synthesis in GA(3)-treated seeds. The rate of incorporation of l-(14) C-leucine into protein was doubled. Autoradiographs showed that enhancement of protein synthesis was localized in the shoot and root meristems, the developing vascular tissues, and in the endosperm cells inside the testa.     

Amino acids in seeds and seedlings of the genus Lens

The amino acid content of seeds and 4-day-old seedlings were studied in five species of lentil: Lens culinaris, L. orientalis, L. ervoides, L. nigricans and L. odemensis. Free amino acid and also total protein amino acid content after HCl hydrolysis were determined by HPLC. The nonprotein UV-absorbing amino acids were determined by capillary zone electrophoresis (CZE). The content of free protein amino acids in seeds varied among species and increased dramatically after germination. Asparagine is quantitatively most important in both seed and seedling. The content of free nonprotein amino acids is variable in seeds and seedlings. gamma-Hydroxyarginine, gamma-hydroxyornithine, alpha-aminobutyric acid and taurine were found in both seeds and seedlings. Homoarginine was found in four species but not in L. orientalis while gamma-aminobutyric acid (GABA), alpha-aminoadipic acid (alpha-aaa) and three isoxazolinone derivatives: beta-(isoxazolin-5-on-2-yl)-alanine (BIA), gamma-glutamyl-BIA (gamma-glu-BIA) and 2-carboxymethyl-isoxazolin-5-one (CMI) were found exclusively in the seedlings. CMI was identified for the first time in lentil species. Lathyrine, beta-(2-amino-pyrimidine-4-yl)-alanine, which was reported to be in the seeds of some Lathyrus species was confirmed to be present also in the seedling of L. culinaris (trace amount), L. nigricans and L. odemensis. Trigonelline (N-methyl-nicotinic acid), a plant hormone, is present both in seeds and seedlings in different concentrations except in L. ervoides. The different combination of nonprotein amino acids among the species gives indication of their genetic relationship and might partly explain the varying compatibility for interspecies crossing.     

Importance of pfkA for rapid growth of Enterobacter cloacae during colonization of crop seeds

Enterobacter cloacae A-11 is a prototrophic, glycolytic mutant of strain 501R3 with a single transposon insertion in pfkA. The populations of strain A-11 on cucumber and radish seeds were smaller than the populations of strain 501R3 in natural soil, but the populations of these two strains on pea, soybean, sunflower, and sweet corn seeds were similar (D. P. Roberts, P. D. Dery, I. Yucel, J. Buyer, M. A. Holtman, and D. Y. Kobayashi, Appl. Environ. Microbiol. 65:2513-2519, 1999). The net effect of the mutation in pfkA in vitro was a shift from rapid growth on certain carbohydrates detected in seed exudates to much slower growth on other carbohydrates, amino acids, and organic acids. The impact of the mutation in pfkA was greatest on the growth rate of E. cloacae on the seeds that released the smallest quantities of fructose, other carbohydrates, and amino acids. Corn, pea, soybean, and sunflower seeds released total amounts of carbohydrates and amino acids at rates that were approximately 10- to 100-fold greater than the rates observed with cucumber and radish seeds for the first 24 h after inhibition began. The growth rate of strain A-11 was significantly less (50% less) than the growth rate of strain 501R3 on radish seeds, and the growth rate of strain A-11 was too low to estimate on cucumber seeds in sterile sand for the first 24 h after inhibition began. The growth rate of strain A-11 was also significantly lower on soybean seeds, but it was only 17% lower than the growth rate of strain 501R3. The growth rates of strains 501R3 and A-11 were similar on pea, sunflower, and corn seeds in sterile sand for the first 30 h after imbibition began. Large reductions in the growth rates of strain A-11 on seeds were correlated with subsequent decreased levels of colonization of seeds compared to the levels of colonization of strain 501R3. The strain A-11 populations were significantly smaller than the strain 501R3 populations only on radish and cucumber seeds. The mutation in pfkA appears to decrease the level of colonization by E. cloacae for seeds that release small quantities of reduced carbon compounds by decreasing the size of the pool of compounds that support rapid growth by this bacterium.