Seed Storage Proteins in Capsicum Annuum Cultivars

Electrophoretic analyses of non-reduced and reduced seed storage proteins from 34 cultivars of Capsicum annuum L. were performed on two gel systems. On the basis of clearly expressed difference in electrophoretic profiles of non-reduced proteins, the investigated cultivars were divided into two groups. The two observed phenotypes were genetically determined and related with intermediary subunits of 11S globulins and/or their oligomers.     

Sequence Peculiarity of Gnetalean Legumin-Like Seed Storage Proteins

The development of seeds as a specialized organ for the nutrition, protection, and dispersal of the next generation was an important step in the evolution of land plants. Seed maturation is accompanied by massive synthesis of storage compounds such as proteins, starch, and lipids. To study the processes of seed storage protein evolution we have partially sequenced storage proteins from maturing seeds of representatives from the gymnosperm genera Gnetum, Ephedra, and Welwitschia—morphologically diverse and unusual taxa that are grouped in most formal systems into the common order Gnetales. Based on partial N-terminal amino acid sequences, oligonucleotide primers were derived and used for PCR amplification and cloning of the corresponding cDNAs. We also describe the structure of the nuclear gene for legumin of Welwitschia mirabilis. This first gnetalean nuclear gene structure contains introns in only two of the four conserved positions previously characterized in other spermatophyte legumin genes. The distinct phylogenetic status of the gnetalean taxa is also reflected in a sequence peculiarity of their legumin genes. A comparative analysis of exon/intron sequences leads to the hypothesis that legumin genes from Gnetales belong to a monophyletic evolutionary branch clearly distinct from that of legumin genes of extant Ginkgoales and Coniferales as well as from all angiosperms. Received: 5 June 1997 / Accepted: 31 March 1998     

水稻种子贮藏谷蛋白的微细异质性

利用灵敏的等聚焦与SDS－PAGE合的双向电泳分析方法,从水稻（Oryza sativa L.)种子贮藏谷蛋白中至少可以分离为13条酸性和19条碱性多 肽,依据谷蛋白多肽的表现量推测,水稻谷蛋白主要由约6个主效基因控制,肽图谱与N－端氨基酸序列分析可清晰将谷蛋白酸性多肽分为两组,此两组恰好与谷蛋白GluA和GluB两个cDNA克隆组相吻合。     

Seed Storage Proteins in Solanaceae and Cucurbitaceae Species

Electrophoretic analyses of non-reduced and reduced seed storage proteins from Solanaceae and Cucurbitaceae species and cultivars were performed. High molecular disulfide bonded complexes between intermediary subunits of 11S globulins previously detected in Capsicum annuum cultivars, were found in Solanum melongena cultivars as well. The data obtained might be used for further elucidation of peculiarities of the 11S globulins in dicotyledonous plants.     

植物种子贮藏蛋白质及其细胞内转运与加工

高等植物种子成熟过程中贮存大量的贮藏蛋白质作为种子发芽和初期生长的重要营养来源。根据溶解性不同,种子贮藏蛋白质可分为白蛋白、球蛋白、醇溶蛋白和谷蛋白4类。在种子胚发育过程中,醇溶蛋白在粗面内质网合成后形成蛋白质聚集体,直接出芽形成蛋白体并贮存其中。白蛋白、球蛋白和谷蛋白在粗面内质网以分子量较大的前体形式合成后,根据各自的分选信号进入特定的运输囊泡,经由受体依赖型运输/聚集体形式运输转运至蛋白质贮藏型液泡中,然后经过液泡加工酶等的剪切转换为成熟型贮藏蛋白质并贮存其中。蛋白质的合成、分选、转运和加工等过程影响种子蛋白质的品质及含量。该文对种子贮藏蛋白质的分类和运输、加工以及这些过程对种子蛋白质品质和含量的影响进行了概述。     

水稻种子贮藏谷蛋白的改良电泳分析法

利用15％－25％丙烯酰胺梯度凝胶的SDS－PAGE分析法可将水稻种子贮藏谷蛋白分离为3个酸性（α）亚基和3个碱性（β）亚基,通过调节两性电解质比例对现有等用点了和焦电泳分析法进行改良,可将谷蛋白酸性亚基和碱性亚基分别分划为13和14条多肽带,将上述两种方法结合起来的双向电泳分析法可以高清晰度地离析谷蛋白并获得单一多肽,此改良的电泳分析系统有助于确定水稻谷蛋白变异及谷蛋白的生化研究。     

Different Types of Major Storage Seed Proteins in Fagaceae Species

The seed proteins of Fagus sylvatica, Quercus Hex and Q. robur,of the Fagaceae family, were extracted by the sequential methodof Koie and Nielsen (1977). The storage proteins of Fagns werefound to be globulins as has previously been shown in Castanea,a member of the same family. Quercus was found to store glutdins.The amino acid composition and the preliminary characterizationby electro-phoresis of Fagus globulins and Quercus glutelins,suggest that both contain proteins similar to the 1 IS globulinsof leguminous plants. Both genera showed considerable differencesin their albumins. Prolamins were found only in Fagus. Key words: Fagaceae, seeds, storage protein     

The Site of Synthesis and Accumulation of Rice Storage Proteins

Electron microscopy showed that the two types of protein bodies(PB) in starchy endosperms of rice were formed differently duringthe period of storage protein accumulation. Two routes for thetransport of storage protein from the site of synthesis at therough endoplasmic reticulum (RER) to the site of accumulationwere also proposed. PB-I, bound by a single membrane to whichribosomes were attached, was thought to develop inside the cisternaeof RER, while the PB-II membrane was thought to originate fromthe vacuole. In the wheat germ cell-free translation system, storage protein-relatedpolypeptides of developing rice endosperms, including a precursorof glutelin and putative precursors of prolamin, were directedby membrane-bound polysomes but not by free-polysomes. Immunoassayof the total translation products directed by a PB fractionshowed that 46% were storage protein-related polypeptides. Rice storage proteins (prolamin) that accumulate in PB-I appearto be synthesized by membrane-bound polysomes attached to PB-Ior RER and to pass through the membrane into the lumen wherethey aggregate and are deposited. The proteins (glutelin andglobulin) that accumulate in PB-II, however, seem to be synthesizedby membrane-bound polysomes as a large precursor and to becomesequestered into the cisternal space of RER, from where theyare transferred to the vacuolar precursor of PB-II. (Received August 6, 1985; Accepted November 6, 1985)     

种子的贮油细胞器——油体及其蛋白

生物柴油作为一种可再生的替代能源,因其良好的环境效应而倍受关注.植物油是生产生物柴油的主要原料.植物油作为最经济、最有利于种子传播和萌发生长的能量和碳源物质,贮存和积累于植物种子的贮油细胞器-油体之中.本文综述了植物种子油体的亚细胞结构、化学组成及其功能、生物合成与降解等的研究进展.     

Promoters of Rice Seed Storage Protein Genes Direct Endosperm-Specific Gene Expression in Transgenic Rice

The promoters of genes encoding rice seed storage proteins (glutelin,prolamin, globulin and albumin) were analyzed for their abilityto direct rß-gIucuronidase (GUS) gene expression intransgenic rice plants. All promoters tested could direct endosperm-specificexpression of the GUS reporter gene irrespective of variableactivities and patterns in the endosperm. (Received February 27, 1998; Accepted May 16, 1998)     

Evolution of the Genes Encoding Seed Storage Proteins in Sugarcane and Maize

Prolamins, the seed storage proteins of maize, sorghum and coix were also found in sugarcane. Prolamins are grouped into structurally distinct classes termed the α-, β-, γ- and δ-prolamins. Orthologues for almost all of the α-, β-, γ- and δ-prolamins classes were identified in sugarcane. In maize, there are two molecular weight classes of α-prolamins, the 22 and 19 kD α-zeins. Sugarcane also possesses both the 22 kD and the 19 kD α-prolamins, which we denote as caneins, whereas sorghum and coix contain only the 22 kD α-prolamin (α-kafirin and α-coixin, respectively). Amino acid sequence alignments of the 22 and 19 kD α-prolamins from these plants revealed that both the 19 kD α-zein and the 19 kD α-canein lack around 20 amino acids at the sixth α-helix domain. We postulate that the 19 kD α-prolamins originated from a deletion of the sixth α-helix of a 22 kD counterpart in the saccharum lineage. Saccharum and sorghum diverged around five to nine million years ago (Mya), when only the 22 kD α-prolamins existed. The 19 kD α-canein must therefore have emerged after this time. Sorghum possesses a 19 kD α-prolamin similar to that of sugarcane and maize, but it contains the sixth α-helix domain lacking in the 19 kD α-zein and the 19 kD α-canein. This sorghum α-prolamin that we called 19 kD-like α-kafirin must be the ancestor of the 19 kD α-canein. The 19 kD-like α-kafirin could also be the ancestor of the 19 kD α-zein but it is also possible that the genes encoding the 19 kD α-zein and the 19 kD α-canein have evolved separately in these close groups.     

RegnANN: Reverse Engineering Gene Networks using Artificial Neural Networks

RegnANN is a novel method for reverse engineering gene networks based on an ensemble of multilayer perceptrons. The algorithm builds a regressor for each gene in the network, estimating its neighborhood independently. The overall network is obtained by joining all the neighborhoods. RegnANN makes no assumptions about the nature of the relationships between the variables, potentially capturing high-order and non linear dependencies between expression patterns. The evaluation focuses on synthetic data mimicking plausible submodules of larger networks and on biological data consisting of submodules of Escherichia coli. We consider Barabasi and Erdös-Rényi topologies together with two methods for data generation. We verify the effect of factors such as network size and amount of data to the accuracy of the inference algorithm. The accuracy scores obtained with RegnANN is methodically compared with the performance of three reference algorithms: ARACNE, CLR and KELLER. Our evaluation indicates that RegnANN compares favorably with the inference methods tested. The robustness of RegnANN, its ability to discover second order correlations and the agreement between results obtained with this new methods on both synthetic and biological data are promising and they stimulate its application to a wider range of problems.     

Isolation and Characterization of Mustard (Sinapis alba L.) Seed Storage Proteins

A total storage protein fraction was prepared from mustard (Sinapis alba L.) seeds via isolated protein bodies and characterized by sedimentation, immunological, and electrophoretic techniques. Mustard seed storage protein consists of three fractions (1) a “legumin-like” 13-S complex composed of two pairs of disulfide-linked polypeptides (16.5 + 28.5 kDa and 19.5 + 34 kDa, respectively) and two single polypeptides (18 kDa and 26 kDa), (2) a “vicilin-like” 9-S complex composed of two glycoproteins (64 kDa and 77 kDa), and (3) two small polypeptides (10 kDa and 11 kDa) which probably represent the 1.7-S complex found in other Cruciferae. In contrast to related species, no glycosylated polypeptide was found in the 13-S complex. Immunological relationships were found between the paired polypeptides of the 13-S complex but not between polypeptides of the 13-S complex and polypeptides of the 9-S complex. Pulse-chase labeling and in vitro translation of polysomal RNA from young embryos demonstrated that the polypeptides of the 13-S complex originate from high molecular mass precursors, except for the 18 kDa polypeptide which appears to be synthesized in its final size. The amino-acid composition of the major polypeptides of the mustard storage protein is given.     

Determination of Seed Storage Proteins and Total Isoflavones in Wild and Cultivated Soybeans

Two soybean components namely, storage proteins and isoflavone content in a wild and three cultivated soybean genotypes were characterized and compared. The storage proteins, β-conglycinin and glycinin were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and two major storage proteins and their subunits were characterized using mass spectrometry. The three isoflavones, aglycon and the nine conjugated forms were separated by HPLC (high performance liquid chromatography) and identified by comparison of retention time, ultraviolet and mass spectral analyses. Comparison between the number of 2D-PAGE protein spots of the storage protein subunits and HPLC area of twelve isoflavones was also evaluated. The analysis of proteins and isoflavones from the wild genotype and the three cultivated genotypes suggested possible interactions between proteins and isoflavones. The same wild genotype, which showed significant statistical differences in β-conglycinin and glycinin protein profiles also revealed considerable reduction in total isoflavones (> 55%) content.     

花生种子发育和萌发过程中贮藏蛋白的合成和降解

以花生品种汕油5 2 3种子为材料,分离纯化花生球蛋白的41 kD和38.5 kD两种主要亚基及伴花生球蛋白的6 0.5 KD亚基并制备抗体.We stern blot分析表明,3种亚基在花生胚组织分化期的胚轴和子叶中就开始合成,其中60.5 kD亚基是最先在胚轴和子叶中大量合成和积累的贮藏蛋白,41 kD和38.5 kD亚基在随后的发育中积累量不断增加;种子萌发时这3种亚基的降解进程不一样,胚轴和子叶中41 kD和38.5kD亚基的降解均先于60.5 kD亚基.     

Comparison of Rice (Oryza sativa L.) Seed Proteins from Several Varieties

Enzymatically modified proteins (EMP) with different methionine levels were produced from soy protein isolate using an improved plastein reaction. The products having methionine at approximate levels of 4%, 7%, and 14%, designated as EMP₄, EMP₇, and EMP₁₄, respectively, were investigated to characterize their chemical properties particularly in terms of the state and location of the methionine residues. Leucine aminopeptidase treatment of the EMP products did not find any significant amount of methionine residues at the N-terminals, but carboxypeptidase A treatment liberated methionine efficiently in accordance with the methionine levels in the EMP products. Treatment with LiBH₄ reduced the methionine content of EMP₁₄ by approximately 64%. A significant amount of homoserine was produced when EMP₁₄ was treated with BrCN. All these data indicate that the covalently attached methionine molecules are localized at or near the C-terminals of the EMP molecules, probably as oligomers.     

Thin Frozen Film Method for Visualization of Storage Proteins in Mature Rice Grains

There are technical difficulties in obtaining intact sections of cereal grains in which mature cells and their subcellular structures are well preserved. Here we describe a simple method for sectioning hard mature rice grains. It makes possible accurate localization of storage proteins in high-quality histological sections of rice endosperm.     

Identification and Characterization of the Seed Storage Proteins from Alfalfa (Medicago sativa)

The major seed storage proteins in alfalfa are medicagin (alegumin-like globulin), alfin (a vicilin-like globulin) anda family of Lower Molecular Weight albumins (LMW₁₃). These comprise30%, 10% and 20%, respectively, of the total extractable proteinfrom cotyledons of mature seeds. Alfin is a heterogeneous oligomericprotein (Mr 150 kD) composed of polypeptides ranging in sizefrom Mr 50 to 14 kD (₁,-₆; 50, 38, 32, 20, 16 and 14 kD, respectively).Medicagin is also a high molecular weight oligomeric protein,but requires high concentrations of salt for solubilization.It is comprised of a family of individually distinct subunits,each composed of an acidic polypeptide (A₁–A₉; Mr 49 to39 kD) linked via disulphide bond(s) to a basic polypeptide(B₁, B₂, B₃; Mr 24, 23 and 20 kD, respectively). This pairingis highly specific and two families are recognizable on thebasis of the B polypeptide (B₃ or B₁/B₂). Subunits (M_r 50–65kD) are assembled as trimers (8S) or larger oligomers (12S–15S)in mature seeds. The lower molecular weight albumins (LMW₁–₃)are acidic (pl<6), and consist of sets of disulphide-bondedpolypeptides (Mr 15 and 11 kD). Key words: Medicago sativa, seed storage proteins, alfin, medicagin