Thesis (M.S.); Submitted to Iowa State Univ., Ames, IA (US); 12 Dec 2003. / Published through the Information Bridge: DOE Scientific and Technical Information. "IS-T 2547" Elizabeth M. Lenihan. 12/12/2003. Report is also available in paper and microfiche from NTIS.

Thesis (Ph. D.)--Miami University, Dept. of Chemistry and Biochemistry, 2008. / Title from second page of PDF document. Includes bibliographical references.

Orientador: Paulo Arruda / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-06T21:19:28Z (GMT). No. of bitstreams: 1 Ferreira_NataliaCristinaVerza_D.pdf: 9495810 bytes, checksum: 18cf9ae055a2b9f39de109abc6de469c (MD5) Previous issue date: 2006 / Resumo: O seqüenciamento de ESTs (etiquetas de seqüências expressas) e a sua organização em bancos de dados constituem poderosas ferramentas para identificar genes de interesse expressos em determinados tecidos e/ou tipos celulares. Neste trabalho criou-se um banco de seqüências expressas chamado MAIZESTdb, que contém ESTs de diversos tecidos de milho, porém enriquecido com seqüências provenientes do endosperma de milho em desenvolvimento. O MAIZESTdb contém 227.431 ESTs vindos de mais de 30 órgãos e tecidos de milho diferentes, 30.531 seqüenciados em nosso laboratório a partir de bibliotecas construídas com RNA mensageiro de endosperma. Estas seqüências representam uma grande contribuição na identificação de novos genes expressos no endosperma. A análise deste banco de ESTs possibilitou a identificação de 4.032 transcritos preferencialmente expressos no endosperma, e a sua anotação revelou uma ampla variedade de prováveis genes novos envolvidos no desenvolvimento e no metabolismo do endosperma. O banco MAIZESTdb foi utilizado neste trabalho para a identificação de fatores de transcrição (TFs) expressos no endosperma de milho, e, especialmente, na identificação de fatores preferencialmente expressos no endosperma, que podem desempenhar papéis regulatórios importantes durante a formação da semente. Foram identificados 1.233 TFs expressos em milho, 414 dos quais expressos no endosperma em desenvolvimento. Foram identificados ainda, através de análises in silico, 113 TFs preferencialmente expressos no endosperma, conjunto este que representa 9.2% dos TFs expressos identificados em milho, e que possivelmente contém reguladores importantes dos processos de especificação celular e desenvolvimento do endosperma de milho. Esta é a maior coleção de fatores de transcrição já descrita para este tecido, e representa uma fonte de dados importante para identificação de reguladores dos principais processos relacionados ao desenvolvimento do endosperma, como metabolismo de nitrogênio e carboidratos e controle da massa da semente. Uma das famílias mais representadas entre os TFs preferencialmente expressos no endosperma foi a família NAC de fatores de transcrição. Esta família apresentou 12 membros preferencialmente expressos no endosperma de milho. Um novo membro da família NAC, chamado de EPN-1 (Endosperm Specific NAM 1), teve seu perfil de expressão caracterizado. Sua expressão pode ser detectada desde os 5 DAPs, embora o pico de expressão ocorra entre 20 e 25 DAP, e ele apresenta expressão preferencial no endosperma. O promotor do gene EPN-1 foi clonado, seqüenciado e analisado quanto aos seus possíveis elementos CIS regulatórios; foram encontrados elementos conservados relacionados à endosperma-especificidade, elementos relacionados à regulação por ácido abscísico e giberelinas, e elementos conservados presentes nos promotores de a-amilases, indicando uma possível relação deste gene com o processo de transição entre a maturação e a germinação da semente. Ensaios de expressão transitória com o promotor do gene EPN-1 revelaram que sua expressão está dirigida à camada de aleurona do endosperma de milho, o que constitui mais uma evidência de sua possível função na regulação de genes relacionados aos processos de maturação e germinação da semente / Abstract: The sequencing of ESTs (expressed sequence tags) and its organization in databases constitute powerful tools to identify genes of interest in certain tissues and/or cell types. In this work we have created MAIZESTdb, a database of ESTs expressed in diverse maize tissues. The importance of this database, however, is that it is enriched with sequences from developing maize endosperm. The MAIZESTdb contains 227,431 ESTs coming from more than 30 different maize tissues and organs, 30,531 of which sequenced from endosperm cDNA libraries constructed in our laboratory. These sequences represent a great contribution for the identification of novel genes expressed in endosperm. The analysis of this ESTs database led to the identification of 4,032 transcripts preferentially expressed in the endosperm, and its annotation revealed a great variety of new genes involved in endosperm metabolism and development. The MAIZESTdb was then used to identify transcription factors (TFs) expressed in maize endosperm, and, mainly, in the identification of TFs preferentially expressed in the endosperm. We identified 1,233 TFs expressed in diverse maize tissues, 414 of which expressed in developing endosperm. We also identified, through in silico comparison of transcript abundance and library source, 113 TFs with preferential expression in endosperm, representing 9,2% of the TFs identified in this work. This dataset probably contains important regulators of cellular specification of the endosperm development. This is the biggest TFs collection reported for this tissue, and represents an important source of data for identification of regulators for main processes related to the endosperm development such as nitrogen and carbohydrate metabolism and control of seed mass. One of the most represented families among the TFs preferentially expressed in endosperm was the NAC family of transcription factors. This family presented 12 members with preferential expression in the endosperm. A new member of the NAC family, called EPN-1 (Endosperm Specific NAM 1), was characterized. Its expression / Doutorado / Genetica Vegetal e Melhoramento / Doutor em Genetica e Biologia Molecular

Transgenic (TG) sorghum genotypes with altered kafirin synthesis were developed by the Africa Biofortified Sorghum Project, employing recombinant DNA technology, with the aim of improving the protein nutritional quality of the grain. In this project, the effects of altered kafirin synthesis on kafirin polymerisation, protein quality, protein body structure and endosperm texture in different TG lines were investigated. The first generation of TG lines were in a type II low-tannin sorghum background. Altered synthesis of different major kafirin sub-classes (α-, γ- and σ-kafirin) was targeted. Some TG lines had improved lysine content (3.17 g/100 g protein) and moderate (55%) to high (74%) cooked in vitro protein digestibility, compared to the parent (2.05 g/100 g protein; 47.4%, respectively). This is of significance as tannins reduce protein digestibility, by complexing with the proline-rich kafirins. Transmission electron microscopy revealed that the improved protein quality traits were associated with floury endosperm texture and irregular protein body structure. Irregular protein bodies were 2-3 ìm in diameter, with few to numerous invaginations, compared to normal protein bodies. The high digestibility TG line also had a unique dense protein matrix, with occasional thick dark-staining inclusions. It appears that reduced kafirin synthesis, specifically γ-kafirin, has a major effect on the protein body structure, which in turn results in changes in protein digestibility and endosperm structure. To further improve the protein quality and poor endosperm texture of the first generation of TG lines, improved non-tannin sorghums were transformed to suppress kafirin synthesis, or they were back-crossed into TG lines with improved protein quality. Co-suppression of the α-, γ- and σ-kafirin sub-classes and removal of the tannin trait, resulted in TG with high cooked protein digestibility (±80%), improved Amino Acid Score (0.8) and Protein Digestibility Corrected Amino Acid Score (0.7) compared to the non-TG null controls (±50%, 0.4 and 0.2, respectively). However, these high-protein quality lines still had a floury endosperm texture. They also had irregular shaped protein body structure, as described previously. When fewer kafirin sub-classes were suppressed (only γ- and σ-kafirin) the endosperm was corneous with normal protein body structure, but the improvement in cooked protein digestibility was less. Apparently, co-suppression of several kafirin sub-classes is required to obtain high-protein quality sorghum, but this seems to result in floury-type grain endosperm. Further work conducted on the high digestible TG line revealed that the proportion of kafirin-1, extracted with 60% tert-butanol alone, was greatly increased. However, the total amount of kafirin remained unchanged. Also, the kafirin was much less polymerised by disulphide bonding, and there was evidence of compensatory synthesis of other kafirin proteins. Hence, the mechanism for the increased protein digestibility of TG lines is probably related to their lower levels of disulphide-bonded kafirins, allowing better access of proteases. This work appears to confirm that disulphide bond formation in kafirin is responsible for the reduced protein digestibility of cooked sorghum. Since grain hardness is an important grain quality attribute, playing a major agronomic role, in sorghum processing and in the end-use quality of sorghum-based foods, further research should focus on transforming sorghum to have both improved protein nutritional quality and good grain endosperm texture. / Thesis (PhD)--University of Pretoria, 2012. / Food Science / unrestricted

The project was carried out in collaboration with Lantmännen with the aim to analyse the lipase activity in oat endosperm, non-heat-treated whole grain oat flour and heat-treated whole grain oat flour and to investigate whether lipase activity could be inactivated in oat endosperm by reducing water activity by various methods such as oven drying and freeze drying to extend oat shelf life. Lipase activity was measured by a wet extraction of the flour and an artificial substrate that gives a coloured product if lipase enzymes are active in the extract. The formation of the product was measured during five minutes at room temperature or 37 °C using a spectrophotometer, and the enzyme activity in units/g of flour was calculated. It was not possible to measure the enzyme activity in the flour directly. The obtained results for the different oat fractions indicated differences in lipase activity between the oat fractions. Lipase activity for oat endosperm was 0.183 U/g, lipase activity for non-heat-treated whole grain oat flour was 0.088 U/mg and the heat-treated whole grain oat flour had no lipase activity. The differences were statistically significant since the p-value was <0.0001. The results from the analysis that compared whether oven-drying and freeze-drying as methods had an effect on lipase activity indicated that oven drying had the greatest effect when assay was performed at room temperature. However, when the assay was performed at 37 °C the lipase activity in oven-dried and freeze-dried oat endosperm were similar to non- treated samples. This indicates that temperature has a great impact on the lipase activity. Thus, oven drying and freeze drying could not completely inactivate the lipase activity, but probably reduce it.

Le grain de maïs est composé de 3 compartiments : l’embryon et l’albumen issus de la double fécondation et l’enveloppe d’origine maternelle. Le développement du grain et l’accumulation de réserves demande l’établissement d’une communication étroite entre l’embryon et l’albumen pour coordonner leur développement respectif. Si, des régulateurs majeurs impliqués dans le développement de la graine d’Arabidopsis ont été décrits, ces connaissances restent parcellaires chez les céréales. Les objectifs de ma thèse consistaient d’une part à étudier le contrôle de la communication entre l’embryon et l’albumen et d’autre part la régulation du remplissage du grain de maïs. Par l’analyse de lignées transgéniques sous exprimant ZmZHOUPI (ZmZOU-RNAi), nous avons établi que ce facteur de transcription à domaine bHLH, bien que s’exprimant exclusivement dans l’albumen, affecte significativement le développement de l’embryon (taille de l’embryon, persistance du suspenseur). L’analyse de données RNAseq (grains sauvages versus grains ZmZOU-RNAi) a permis d’identifier des gènes cibles potentiels de ZmZOU. De plus, nous avons montré que 3 facteurs de transcription de type bHLH homologues d’INDUCER OF CBP EXPRESSION (ICE) forment un partenariat avec ZmZOU.D’autre part, nous avons étudié les homologues d'ABA INSENSITIVE3, FUSCA3 et LEAFY COTYLEDON2 (AFL) qui forment un réseau de facteurs de transcription, à domaine B3, régulant l’accumulation d’huile et de protéines de réserves dans l’embryon d’Arabidopsis. Grâce à des analyses phylogénétiques et d’expression, nous avons établi que chez le maïs le réseau AFL, constitué de 5 membres (ZmAFLs), est partiellement conservé. Par dosages et analyse d’expression, nous avons montré que ZmAFL4, en particulier, est impliqué dans le contrôle de la biosynthèse de l’amidon dans l’albumen. / Maize kernel is composed of three major compartments: an embryo and an endosperm both produced by double fertilization and the maternally derived seed coat. Seed development and reserves accumulation demands coordination and thus communication between embryo and endosperm allowing specific growth of each compartment. While major regulators involved in seed development have been already described in Arabidopsis, knowledge in cereals remains limited. My thesis purposes were to study on one hand the control of communication between embryo and endosperm and on the other hand regulation of maize kernel filling.By analysis of transgenic lines knock down ZmZHOUPI (ZmZou-RNAi), we showed that this bHLH domain transcription factor, exclusively expressed in endosperm, affect significantly embryo development, size of embryo proper and suspensor persistence. RNAseq data analyses let find putative direct targets of ZmZOU. Additionally, we identified ZmZOU partners, 3 bHLH domain transcription factor homologs of INDUCER OF CBP EXPRESSION (ICE).Furthermore, we studied homologs of three B3 domain transcription factors named ABA INSENSITIVE3, FUSCA3 et LEAFY COTYLEDON2 (AFL) which form a regulatory network governing oil and seed storage proteins accumulation in Arabidopsis embryo. By phylogenetic and expression analysis, we established that 5 genes (ZmAFLs) constitute in maize a partially conserved AFL network. Through dosage and expression analysis, we established that particularly ZmAFL4 is involved in starch biosynthesis regulation.

Thesis (MScAgric (Animal Sciences))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: The purpose of this study was to investigate the variation that exists between maize samples regarding particle size separation, in vitro fermentation kinetics and in vitro dry matter (DM) disappearance. A second objective was to quantify possible relationships between the Roff Milling Index (RMI) of maize and any of the measured in vitro parameters. Three trials were conducted: a particle size distribution trial, a gas production trial and an in vitro DM degradability and starch disappearance trial. Overall, nine maize samples, which differed in terms of cultivar and endosperm type, were collected from different origins for the study. The samples were selected in terms of their Milling Index (MI). Three of the nine samples had a high MI that ranged between 109 and 118, three had a low MI that ranged between 67 and 71 and the other three samples had a medium MI that ranged between 85 and 92. Although the MI is not a direct indication of the hardness or softness of the endosperm, it was believed to be indirectly associated with vitreousness. In the first trial, the different maize samples were milled through a 1 mm screen and sieved through a series of three sieves 150, 125 and 106 μm, respectively. It was found that RMI was not a reliable indicator to predict particle size distribution, especially in terms of the coarse (>150 μm) and very fine (<106 μm) particles. In the gas production trial, the nine different maize samples were subjected to a gas production system for a duration of 48 hours. Here gas production and rate of gas production of the different maize types in buffered rumen liquor were measured during incubation. After fitting the gas volume data to the respective models, the non-linear parameters b, c and L were subjected to a main effects ANOVA with the aid of Statistica, version 9 (2009). Main effects were treatment and repetition. Means were separated by means of a Scheffé test and significance was declared at P < 0.05. The results were compared to the RMI of the different maize types and it was concluded that RMI was not a reliable predictor of gas production or rate of gas production of different maize types. In the third trial, in vitro DM degradability and starch disappearance of the different maize types were measured. In vitro DM degradability was conducted in the Ankom DAISYII incubator apparatus and the incubation times were 0, 2, 4, 8, 12 and 24 hours. Starch disappearance was measured on residues of the samples incubated for 0, 2 and 4 hours. After fitting the DM disappearance data to the respective models, the non-linear parameters a, b, c and L were subjected to a main effects ANOVA with the aid of Statistica, version 9 (2009). Main effects were treatment and repetition. Means were separated by means of a Scheffé test and significance was declared at P<0.05. The results indicated variation between maize samples, especially in terms of the a-, b- and L-values. The RMI did not appear to be a reliable predictor of digestibility parameters. / AFRIKAANSE OPSOMMING: Die doel van hierdie studie was om die variasie tussen mieliemonsters te ondersoek ten opsigte van die skeiding van partikelgroottes, in vitro-fermentasiekinetika en in vitro-droëmateriaalverdwyning. ‘n Tweede doel was om te bepaal of daar moontlike verwantskappe tussen die Roff Milling Index (RMI) van mielies en enige van die ander in vitro-parameters bestaan. Drie proewe is gedoen: verspreiding van partikelgrootte, ‘n gasproduksieproef en ‘n droëmateriaal degradeerbaarheid- en stysel verdwyningsproef. Nege mieliemonsters, wat van mekaar verskil ten opsigte van kultivar en endospermtipe, is van verskillende lokaliteite versamel. Die monsters is gekies in terme van hul maal-indeks (MI). Drie van die nege monsters het ‘n hoë MI gehad wat gewissel het tussen 109 en 118, drie het ‘n lae MI gehad wat gewissel het tussen 67 en 71 en die ander drie monsters het ‘n medium MI gehad wat gewissel het tussen 85 en 92. Alhoewel die MI waardes nie ‘n direkte indikasie van ‘n endosperm se hardheid- of sagtheidsgraad is nie, is dit aanvaar dat daar ‘n indirekte verwantskap tussen MI en glasagtigheid van die mielie bestaan. In die eerste proef is die nege verskillende mieliemonsters deur ‘n 1 mm sif gemaal en daarna deur ‘n reeks van drie siwwe met groottes van onderskeidelik 150, 125 en 106 μm gesif. Daar is bevind dat die RMI nie ‘n betroubare voorspeller is om partikelgrootte-verspreiding aan te dui nie, veral nie ten opsigte van growwe (> 150 μm) en baie fyn (< 106 μm) patikels nie. Tydens die gasproduksieproef is die nege mieliemonsters vir 48 ure blootgestel aan ‘n gasproduksiesisteem, waar gasdruk outomaties aangeteken is. Gasproduksie en tempo van gasproduksie van die verskillende mieliemonsters is gemeet en aangeteken gedurende inkubasie met ‘n gebufferde rumenvloeistofmedium. Nadat die gasvolumedata met behulp van relevante modelle gepas is, is die nie-linêre parameters b, c en L onderwerp aan ‘n hoof-effek ANOVA met die gebruik van Statistica weergawe 9 (2009). Hoof-effekte was behandeling en herhaling. Gemiddeldes is deur ‘n Scheffé-toets geskei en betekenisvolheid is verklaar by P<0.05. Die resultate verkry is vergelyk met die RMI van die verskillende mielietipes. Die gevolgtrekking is gemaak dat Roff MI nie ‘n betroubare voorspeller van totale gasproduksie of gasproduksietempo is nie. Tydens die derde proef is droëmateriaaldegradeerbaarheid en styselverdwyning van die verskillende mielietipes bepaal. In vitro droëmateriaal (DM) degradeerbaarheid is gedoen in die Ankom DAISYII inkubator met inkubasietye van 0, 2, 4, 8, 12 en 24 ure. Styselverdwyning is bepaal deur styselanalises op die residue van die monsters wat geïnkubeer is vir 0, 2 en 4 ure. Nadat die DM-degradeerbaarheid met behulp van relevante modelle gepas is, is die nie-lineêre parameters a, b, c en L onderwerp aan ‘n hoof-effek ANOVA met die gebruik van Statistica weergawe 9 (2009). Hoof-effekte was behandeling en herhaling. Gemiddeldes is deur ‘n Scheffé toets geskei en die betekenisvolheid is verklaar by P<0.05. Die resultate het aangedui dat daar groot variasie tussen mielies bestaan, veral ten opsigte van die a-, b- en L-waardes. Dit het verder geblyk dat die RMI van die verskillende mielietipes nie ‘n betroubare voorspeller van DM-degradeerbaarheid was nie.

Chez la plante modèle Arabidopsis, le processus de maturation de la graine et, en particulier, l’accumulation de composés de réserves (huile et protéines de réserve) sont étudiés depuis de nombreuses années. Si les voies de biosynthèse conduisant à l’accumulation de tels composés sont bien décrites, leur régulation est encore largement méconnue. Mon travail de thèse s’inscrit dans un projet de recherche dont le but est d’identifier de nouveaux régulateurs transcriptionnels de la maturation de la graine d’Arabidopsis. Après avoir réalisé une étude comparative du processus de maturation chez les deux zygotes de la graine, embryon et albumen, nous avons caractérisé un facteur de transcription appelé MYB118, exprimé spécifiquement dans l’albumen et potentiellement impliqué dans la régulation du processus de maturation. Son patron d’expression, finement caractérisé, montre un pic d’accumulation d’ARNm en début de maturation de la graine, plus spécifiquement dans l’albumen. Des études menées sur des lignées mutantes ou surexprimant LEAFY COTYELDON2 (LEC2) révèlent que l’expression de MYB118 est activée par ce régulateur maître de la maturation de la graine. Une analyse biochimique de graines myb118 montre que le contenu en huile et en protéines de réserve est doublé dans l’albumen et réduit dans l’embryon de ce mutant par comparaison aux graines sauvages. Finalement, une analyse transcriptomique effectuée sur des graines myb118 a permis d’identifier des cibles putatives dont la dérégulation pourrait expliquer le phénotype : MYB118 semble être un répresseur de l’accumulation de composés de réserve dans l’albumen. Comme la famille de facteurs de transcription à laquelle appartient MYB118 comprend de nombreux membres, nous nous sommes intéressés au patron d’expression et au rôle de ses paralogues les plus proches. L’un d’entre eux, appelé MYB115, est exprimé spécifiquement dans l’albumen chalazal et semble avoir une fonction partiellement redondante à celle de MYB118. / In the model plant Arabidopsis thaliana, seed maturation and more especially the accumulation of storage compounds such as oil and seed storage proteins (SSP) have been widely studied. Although the biosynthetic networks underlying the accumulation of such compounds are now well described, regulation of these pathways remains poorly understood. My Ph.D. project is a part of a research program aimed at identifying new transcriptional regulators of seed maturation in Arabidopsis. After a comparative analysis of maturation processes in the two zygotic tissues of the seed, namely the embryo and the endosperm, we have characterized MYB118, an endosperm-specific transcription factor, putatively involved in the regulation of this maturation process. A fine and comprehensive characterization of its expression pattern showed a peak of expression at the onset of the maturation phase in the endosperm. Expression studies carried out in LEAFY COTYLEDON2 (LEC2) mutant and over-expressing lines demonstrated that MYB118 expression is positively regulated by this master regulator. Biochemical analysis of myb118 seeds showed that oil and SSP contents were doubled in the endosperm fraction and decreased in the embryo of this mutant compared to the wild type. A transcriptomic analysis of myb118 mutant seeds point out some putative targets, the misregulation of which could explain this phenotype: MYB118 seems to be a repressor of storage compounds accumulation in the endosperm.Since MYB118 belongs to the broad family of MYB transcription factors, we investigated the expression pattern and the role of its closest paralogs. One of them, called MYB115 is expressed specifically in the chalazal endosperm and seems to have partially redundant functions with MYB118.

A principal fonte de proteínas para alimentação humana e animal é fornecida pelas sementes de cereais e leguminosas. O conteúdo de aminoácidos solúveis em endospermas de milho normal e mutantes opaco-2 e floury foram determinadas por HPLC. A análise indicou que a concentração total de aminoácidos solúveis variou entre os mutantes e seus tipos selvagens. Nos mutantes o10, o11 e o13, as concentrações foram aumentadas significativamente quando comparadas ao tipo selvagem W22, enquanto os mutantes o1, o2, o13, fl1 e fl2 exibiram baixas concentrações em relação ao seu respectivo tipo selvagem Oh43. Resultados similares foram obtidos para os mutantes o5, o7 e fl3 em relação aos seus tipos selvagens (B79, B37 e WT3, respectivamente). Para metionina, o mutante o2 e o tipo selvagem Oh43 apresentaram as mais altas concentrações deste aminoácido. Diferenças significativas não foram observadas para os outros aminoácidos analisados, tais como lisina e treonina. Os resultados sugerem que as altas concentrações sugeridas originalmente para estes mutantes devem ser devidas aos níveis destes aminoácidos incorporados nas proteínas de reserva, mas não na forma solúvel. / For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperms of wildtype and maize opaque and floury mutants have been determined by HPLC. The total absolute concentration of soluble amino acids among the mutants and their wild-type counterparts varied depending on the mutant. In the o10, o11 and o13 mutants the concentrations were significantly increased when compared to their wild-type counterpart W22, whereas the mutants o1, o2, o13, fl1 and fl2 exhibited lower concentrations when compared to the wild-type Oh43, Similar results were observed for o5, o7 and fl3 in relation to their specific wild-type counterparts (B79, B37 and WT3, respectively). For soluble methionine content, o2 and Oh43 exhibited the highest concentrations. Significant differences were not observed for other amino acids such as lysine and threonine. The results suggest that the high-lysine concentrations indicated originally for these mutants must be due to the amino acids incorporated into storage proteins, but not in the soluble form.

A principal fonte de proteínas para alimentação humana e animal é fornecida pelas sementes de cereais e leguminosas. O conteúdo de aminoácidos solúveis em endospermas de milho normal e mutantes opaco-2 e floury foram determinadas por HPLC. A análise indicou que a concentração total de aminoácidos solúveis variou entre os mutantes e seus tipos selvagens. Nos mutantes o10, o11 e o13, as concentrações foram aumentadas significativamente quando comparadas ao tipo selvagem W22, enquanto os mutantes o1, o2, o13, fl1 e fl2 exibiram baixas concentrações em relação ao seu respectivo tipo selvagem Oh43. Resultados similares foram obtidos para os mutantes o5, o7 e fl3 em relação aos seus tipos selvagens (B79, B37 e WT3, respectivamente). Para metionina, o mutante o2 e o tipo selvagem Oh43 apresentaram as mais altas concentrações deste aminoácido. Diferenças significativas não foram observadas para os outros aminoácidos analisados, tais como lisina e treonina. Os resultados sugerem que as altas concentrações sugeridas originalmente para estes mutantes devem ser devidas aos níveis destes aminoácidos incorporados nas proteínas de reserva, mas não na forma solúvel. / For human nutrition the main source of vegetable proteins are cereal and legume seeds. The content of total soluble amino acids in mature endosperms of wildtype and maize opaque and floury mutants have been determined by HPLC. The total absolute concentration of soluble amino acids among the mutants and their wild-type counterparts varied depending on the mutant. In the o10, o11 and o13 mutants the concentrations were significantly increased when compared to their wild-type counterpart W22, whereas the mutants o1, o2, o13, fl1 and fl2 exhibited lower concentrations when compared to the wild-type Oh43, Similar results were observed for o5, o7 and fl3 in relation to their specific wild-type counterparts (B79, B37 and WT3, respectively). For soluble methionine content, o2 and Oh43 exhibited the highest concentrations. Significant differences were not observed for other amino acids such as lysine and threonine. The results suggest that the high-lysine concentrations indicated originally for these mutants must be due to the amino acids incorporated into storage proteins, but not in the soluble form.