Advances in Tissue Culture and Transformation Studies in Non-model Species: Passiflora spp. (Passifloraceae).

In this chapter, we report advances in tissue culture applied to Passiflora. We present reproducible protocols for somatic embryogenesis, endosperm-derived triploid production, and genetic transformation for such species knowledge generated by our research team and collaborators in the last 20 years. Our research group has pioneered the work on passion fruit somatic embryogenesis, and we directed efforts to characterize several aspects of this morphogenic pathway. Furthermore, we expanded the possibilities of understanding the molecular mechanism related to developmental phase transitions of Passiflora edulis Sims. and P. cincinnata Mast., and a transformation protocol is presented for the overexpression of microRNA156.

Histogenesis and reserve dynamics during the maintenance of dormancy and germination in seeds of the basal palm Mauritiella armata.

The germination and post-seminal development of Arecaceae are notably complex due to the microscopic dimensions of the embryonic axis, the occurrence of dormancy, and the diversity of reserve compounds. In-depth information on this subject is still limited, especially in terms of the basal sub-family Calamoideae. Mauritiella armata is widely distributed in the Amazon region and is considered a key species in flooded ecosystems (veredas) in the Cerrado biome. We sought to describe histogenesis and reserve compound dynamics during the germination of M. armata, as well as the changes in incubated seeds over time. Seeds with their operculum removed (the structure that limits embryonic growth) were evaluated during germination using standard methods of histology, histochemistry, and electron microscopy. Evaluations were also performed on intact seeds incubated for 180 days. The embryos show characteristics associated with recalcitrant seeds of Arecaceae: a high water content (>80%), differentiated vessel elements, and reduced lipid reserves. Both the embryo and endosperm store abundant reserves of proteins, neutral carbohydrates, and pectins. The completion of germination involves cell divisions and expansions in specific regions of the embryo, in addition to the mobilization of embryonic and endospermic reserves through symplastic and apoplastic flows. Intact seeds show dormancy (not germinating for 180 days), but exhibit continuous development associated with cell growth, differentiation, and reserve mobilization. The anatomical and histochemical characters of M. armata seeds indicate an association between recalcitrance and dormancy related to the species' adaptation to flooded environments.

Proteome Landscape during Ripening of Solid Endosperm from Two Different Coconut Cultivars Reveals Contrasting Carbohydrate and Fatty Acid Metabolic Pathway Modulation.

Cocos nucifera L. is a crop grown in the humid tropics. It is grouped into two classes of varieties: dwarf and tall; regardless of the variety, the endosperm of the coconut accumulates carbohydrates in the early stages of maturation and fatty acids in the later stages, although the biochemical factors that determine such behavior remain unknown. We used tandem mass tagging with synchronous precursor selection (TMT-SPS-MS3) to analyze the proteomes of solid endosperms from Yucatan green dwarf (YGD) and Mexican pacific tall (MPT) coconut cultivars. The analysis was conducted at immature, intermediate, and mature development stages to better understand the regulation of carbohydrate and lipid metabolisms. Proteomic analyses showed 244 proteins in YGD and 347 in MPT; from these, 155 proteins were shared between both cultivars. Furthermore, the proteomes related to glycolysis, photosynthesis, and gluconeogenesis, and those associated with the biosynthesis and elongation of fatty acids, were up-accumulated in the solid endosperm of MPT, while in YGD, they were down-accumulated. These results support that carbohydrate and fatty acid metabolisms differ among the developmental stages of the solid endosperm and between the dwarf and tall cultivars. This is the first proteomics study comparing different stages of maturity in two contrasting coconut cultivars and may help in understanding the maturity process in other palms.

A two-year study on the occurrence and concentration of mycotoxins in corn varieties with different endosperm textures.

BACKGROUND: Mycotoxin monitoring in cereal grains has great importance in the food and feed industries. This study evaluated mycotoxin contamination in corns with different endosperm textures in 2 years of cultivation. Samples of dent, semi-dent, flint and semi-flint corns from field experiments were analyzed by high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). RESULTS: Occurrences of fumonisins B1 (FB1 ) and B2 (FB2 ) in 2020 were 45.72% (mean 270 µg kg-1 ) and 35.89% (94.97 µg kg-1 ), respectively, and 68.98% (446 µg kg-1 ) and 45.83% (152 µg kg-1 ) in 2021. Occurrence of aflatoxin B1 was 11.96% (0.16 µg kg-1 ) in 2020 and 11.11% (0.13 µg kg-1 ) in 2021. In 2020, deoxynivalenol (DON) and zearalenona (ZEA) presented occurrences of 1.28% and 1.70%, with means of 4.08 and 2.45 µg kg-1 , respectively. In 2021, results were 8.33% (31.00 µg kg-1 ) for DON and 8.79% (4.38 µg kg-1 ) for ZEA. Citrinin, diacetoxyscirpenol and fusarenon-X did not occur in 2020 but presented 1.66%, 0.83%, and 2.50% positive rates in 2021, respectively. In 2020, flint corn presented the lowest concentration of FB1 whereas dent corn presented the highest concentration of FB1 and FB2 (P < 0.05). In 2021, dent corn presented the highest means of FB1 , FB2 and diacetoxyscirpenol (P < 0.05). Dent and semi-dent presented the highest concentration of nivalenol (P < 0.05). CONCLUSION: The endosperm texture influenced mycotoxin contamination in corn grains, especially FB1 and FB2 , which had the highest concentration in dent corn in the 2 years of this study. © 2023 Society of Chemical Industry.

The effect of the electromagnetic field on the physicochemical properties of isolated corn starch obtained of plants from irradiate seeds.

Maize grains are composed of the pericarp, endosperm, and germ. Consequently, any treatment, such as electromagnetic fields (EMF) must alter these components, which in turn alters the physicochemical properties of the grain. Since starch is a major component of corn grain, and given the great industrial importance of starch, this study investigates how EMF affects the physicochemical properties of starch. Mother seed were exposed to three different intensities 23, 70, and 118 µT for 15 days. Except for a slight porosity on the surface of the starch of the grains of plants exposed to higher EMF, the starch showed no morphological differences between the different treatments and the control (according to scanning electron microscopy). The X-ray patterns showed that the orthorhombic structure was kept constant, unaffected by the intensity of EMF. However, the pasting profile of starch was affected, and a decrease in the peak viscosity was obtained when the intensity of EMF increased. In contrast to the control plants, FTIR shows characteristic bands which can be attributed to the stretching of the CO bonds at wave number 1.711 cm-1. EMF can be considered a physical modification of starch.

CRYPTOCHROME 1a-mediated blue light perception regulates tomato seed germination via changes in hormonal balance and endosperm-degrading hydrolase dynamics.

MAIN CONCLUSION: Blue light exposure delays tomato seed germination by decreasing endosperm-degrading hydrolase activities, a process regulated by CRY1a-dependent signaling and the hormonal balance between ABA and GA. The germination of tomato seeds (Solanum lycopersicum L.) is tightly controlled by an internal hormonal balance, which is also influenced by environmental factors such as light. In this study, we investigated the blue light (BL)-mediated impacts on physiological, biochemical, and molecular processes during the germination of the blue light photoreceptor CRYPTOCHROME 1a loss-of-function mutant (cry1a) and of the hormonal tomato mutants notabilis (not, deficient in ABA) and procera (pro, displaying a GA-constitutive response). Seeds were germinated in a controlled chamber in the dark and under different intensities of continuous BL (ranging from 1 to 25 µmol m-2 s-1). In general, exposure to BL delayed tomato seed germination in a fluency rate-dependent way due to negative impacts on the activities of endosperm-degrading hydrolases, such as endo-ß-mannanase, ß-mannosidase, and α-galactosidase. However, not and pro mutants presented higher germination speed index (GSI) compared to WT despite the BL influence, associated with higher hydrolase activities, especially evident in pro, indicating that the ABA/GA hormonal balance is important to diminish BL inhibition over tomato germination. The cry1a germination percentage was higher than in WT in the dark but its GSI was lower under BL exposure, suggesting that functional CRY1a is required for BL-dependent germination. BL inhibits the expression of GA-biosynthetic genes, and induces GA-deactivating and ABA-biosynthetic genes. The magnitude of the BL influence over the hormone-related transcriptional profile is also dependent upon CRY1a, highlighting the complex interplay between light and hormonal pathways. These results contribute to a better understanding of BL-induced events behind the photoregulation of tomato seed germination.

Ananas ananassoides (Baker) L.B.Sm. a bromeliad from the savanna: seed morpho-anatomy and histochemistry.

Ananas ananassoides (Baker) L.B.Sm. is a wild pineapple, commonly found in the savannas. This study aimed to describe the morpho-anatomy and histochemistry of its seed. The observations were made in the longitudinal and transverse sections, using an optical microscope. The cell arrangement in the seed coat, ripples in the integument, the ratio of embryo size and endosperm amount, and the number of strata in the aleurone layer are anatomical characteristics that may contribute to distinguishing this species. The starch in the endosperm, lipids and proteins in the embryo, constitute the seed's main nutritional reserves. The homogeneous embryo and phenolic compounds present in the seed coat and in the aleurone layer possibly contribute to the dormancy in this species. This study presents information relevant to the taxonomy and physiology of A. ananassoides, which represents contributions to the global knowledge of this species with a high potential as ornamental.

Maize dispersal patterns associated with different types of endosperm and migration of indigenous groups in lowland South America.

BACKGROUND AND AIMS: The lowlands of South America appear to be remarkably important in the evolutionary history of maize, due to new evidence that suggests that maize dispersed from Mexico and arrived in this region in a state of partial domestication. This study aimed to identify dispersal patterns of maize genetic diversity in this part of the continent. METHODS: A total of 170 maize accessions were characterized with 4398 single nucleotide polymorphisms (SNPs) and analysed to determine if maize dispersal was associated with types of endosperm and indigenous language families. KEY RESULTS: Four genetic groups were identified in the discriminant analysis of principal components and five groups in the cluster analysis (neighbour-joining method). The groups were structured according to the predominance of endosperm types (popcorn, floury, flint/semi-flint). Spatial principal component analysis of genetic variation suggests different dispersal patterns for each endosperm type and can be associated with hypotheses of expansions of different indigenous groups. CONCLUSIONS: From a possible origin in Southwestern Amazonia, different maize dispersal routes emerged: (1) towards Northern Amazonia, which continued towards the Caatinga and south-eastern Atlantic Forest (Floury); (2) towards Southern Brazil, passing through the Cerrado and Southern Atlantic Forest reaching the Pampa region (Floury); and (3) along the Atlantic Coast, following Tupi movements originating from two separate expansions: one (Tupinamba) from north to south, and the other (Guarani) in the opposite direction, from south to north (flint, floury and popcorn).

Environment affects starch composition and kernel hardness in temperate maize.

BACKGROUND: Protein percentage and kernel weight affect the endosperm hardness of maize grains. However, changes in starch composition could also modify kernel hardness, which is often predicted through milling ratio. The objective of this work was to evaluate the relationship between changes in starch composition and endosperm hardness, and to assess the effects of protein content and kernel weight on that relationship. For this, we used information obtained from three temperate hybrids in multi-environmental experiments, as well as experiments conducted under controlled conditions designed to modify starch composition. Milling ratio was determined as maize kernel hardness predictor in both experiments and it was correlated with grain weight and grain composition. RESULTS: In both experiments, milling ratio presented a significant Spearman correlation coefficient with amylose/starch ratio. Milling ratio was significantly related to the amylose/starch ratio using a simple fit with datasets from different experiments and hybrids. Increases in amylose/starch ratio were associated with increases in milling ratio. CONCLUSION: Starch composition was related to milling ratio variations among hybrids of different aptitude for dry milling through different environments, regardless of protein content. Thus, increases in the amylose/starch ratio were related to increases in the milling ratio. © 2022 Society of Chemical Industry.

Spatio-temporal characterization of the fruit metabolism in contrasting accessions of Macauba (Acrocomia aculeata).

Although Macauba (Acrocomia aculeata) has been highlighted by its high-quality oil to fit edible and nonedible purposes, data addressing carbon and nitrogen metabolism underlying development and ripening of fruits remain scarce. In addition, accessions of Macauba exibit varied oil yield in fruits, including during the fruit development stages. Here, we monitored contents of carbohydrates, proteins, amino acids and lipids in the mesocarp and endosperm of Macauba fruits until ripening. We selected three accessions from different Brazilian regions (southeast, MG; northeast, PE; and central-west, MS) that differ in the mesocarp lipid content of ripe fruits. Despite the anatomical differences, mesocarp and endosperm exhibited similar trends of metabolite accumulation for most of the analyzed compounds. In the mesocarp, total soluble protein, free amino acids, sucrose, starch and total lipids accumulate towards ripening, while glucose and fructose declined in all accessions. Endosperm differed from mesocarp solely in the amino acid content, which decreased in ripe fruits. In the endosperm, accessions accumulated carbohydrates differently. Accession PE showed comparable fructose and starch contents in the endosperm between the beginning of fruit development and ripening, while in accessions MG and MS, both compounds decreased and increased, respectively, towards ripening. Accession MG was highlighted by its highest lipid content in the two tissues indicating its potential for energy and cosmetic industries. Our results provide novel insights into metabolic changes underlying development and ripening of Macauba fruits and variability in oil content among accessions, indicating new targets for breeding programs.

A comparative approach to understanding the ovule, seed, and fruit development in Bulbostylis (Cyperaceae: Cyperoideae: Abildgaardieae).

In the present work, we study the ovule, seed, and fruit development in six Bulbostylis species in order to characterize the genus in a comparative approach and to identify the characteristics that can be used in taxonomy and phylogeny. Flowers and fruits at different developmental stages were analyzed using LM and SEM after processing according to standard techniques. The species studied have the following: anatropous and bitegmic ovules, weak crassinucellar ovules, obturator of integumentary origin, monosporic embryo sac of the Polygonum type, nuclear endosperm, hypostase formation, seed coat formed by tanniferous endotegmen and exotesta, and Bulbostylis-type embryo. On the other hand, the pericarp development constitutes the main variation within Bulbostylis since the cells of the exocarp may or may not present starch grains, and their inner periclinal walls may be slightly or deeply concave depending on the degree of development of the mesocarp sclereids. In a taxonomic context, the results herein obtained are in conflict with studies which suggest infrageneric groupings based on fruit micromorphology, and also with the relationship among the Bulbostylis species based on molecular analysis. This work contributes to a better understanding of the reproductive anatomy and embryology in Bulbostylis, and reveals the first insights about the origin of multiple embryos in Cyperaceae. Given the frequent presence of polyembryony in Bulbostylis, and the poor mention of this condition in the family, this work highlights an aspect in the anatomy of Cyperaceae that must be re-explored.

Kernel characterization and starch morphology in five varieties of Peruvian Andean maize.

Peruvian Andean maize (PAM) has been commonly used as an ingredient that confers color, flavor, and texture in culinary. Nevertheless, no studies are focusing on agro-industrial interest characteristics to develop new products. This study aimed to evaluate the physicochemical, nutritional, and technological characteristics of kernels and the starch granule morphology of the five main PAM varieties: Chullpi, Piscorunto, Giant Cuzco, Sacsa, and Purple. PAM's characterization was performed according to the official methods, and its morphology was observed by scanning electron microscopy (SEM). Physically, the varieties of larger kernels (Giant Cuzco and Sacsa) presented a higher 1000-kernel weight and a lower hectoliter weight than those of smaller size (Piscorunto, Purple, and Chullpi). Nutritionally, PAM had higher ether extract (5%) and ash (2%) contents than other pigmented maizes. Likewise, they presented more significant amounts of essential amino acids, as leucine (10 mg/g protein) and tryptophan (up to 2 mg/g protein); unsaturated fatty acids, oleic (30%) and linoleic (53%); and minerals, as magnesium (104 mg/100 g). SEM showed that endosperm structure and starch morphology vary according to maize types and their grain location. Starch granules of floury PAM varieties were small and polyhedral in the sub-aleurone endosperm, whereas those of the central area were bigger and spherical. In Chullpi, it was observed a portion of vitreous endosperm with a compact structure. The low protein content (8.3%) and the endosperm structure of floury varieties of PAM influenced their pasting properties. Their pasting temperature was <69 to 71 °C>, peak viscosity < 3200 to 4400 cP>, and seatback <1250 to 1706 cP>; therefore, they do not retrograde easily. The results suggest that PAM has characteristics that would help elaborate regional products with added value, such as soups, willows, beverages, and porridges.

Five-year cryopreservation at -80 °C of edible and medicinal basidiomycetes by wheat grain technique.

This research has focused on basidiomycete cryopreservation at -80 °C and developed a cryopreservation method based on the use of hard or medium-hard endosperm wheat grains as a mycelial carrier for cryopreservation. The aim of this study was to evaluate the mycelial viability of edible and medicinal basidiomycetes, using 13 strains of Agaricus spp. and eight strains of non-Agaricus spp., cryopreserved at -80 °C on hard endosperm wheat grain, with or without cryoprotectant agent (4% glucose), for two and five years. Two groups of basidiomycetes, Agaricus genus and other non-Agaricus genera, were cryopreserved at -80 °C by wheat grain technique for two and five years. The cryopreservation technique with hard endosperm wheat grain without cryoprotectant (preservation substrate), settled previously for A. subrufescens is efficient to cryopreserve other basidiomycetes such as Lentinus crinitus, Pleurotus ostreatus, Pleurotus eryngii, Schizophyllum commune, and Lentinula edodes, besides A. subrufescens strains.

Proteome dynamics of the cotyledonary haustorium and endosperm in the course of germination of Euterpe oleracea seeds.

The role of the cotyledonary haustorium (CH) in the mobilization of nutrient reserves in the endosperm of species of the palm family Arecaceae is a moot question. To shed light on this matter, we present here an analysis of the quantitative proteome changes associated with four developmental stages of CH and three of endosperm during germination. Together, a total of 1965 proteins were identified, being 1538 in the CH and 960 in the endosperm. Both in the CH and endosperm proteomes, we observed an increase in the diversity of hydrolases as the CH and endosperm develops. Qualitative proteomics analysis of four CH developmental stages indicated that each stage is populated by a unique set of proteins and the quantitative analysis showed an increase in the relative abundance of hydrolases, particularly mannan degrading enzymes, as development progresses. These results add weight to the hypothesis that the CH in the seeds of E. oleraceaacts both as a conduit of carbon and nitrogen sources generated by the hydrolysis of the reserves in the endosperm and as a source of hydrolases that will contribute to the mobilization of these reserves.

Amylose content and micromorphology of popcorn progenies with different popping expansion volumes / Teores de amilose e micromorfologia de progênies de milho-pipoca com diferentes volumes de expansão

Popcorn (Zea mays var. everta) has a higher commercial value than common maize, in addition to being a popular food among consumers. Today, there is a constant search for cultivars with superior performance for several traits of interest in the case of popcorn, yield and popping expansion. On this basis, this project proposes to characterize progenies of popcorn with different values ​​of expansion capacity regarding chemical composition and micromorphology. Kernels from the fifth cycle (C5) of intrapopulation recurrent selection were evaluated. The progenies were selected based on the popping expansion volume of their kernels. The kernels were quantified for amylose and analyzed for starch granule arrangement and pericarp thickness by scanning electron microscopy. Progenies with low popping expansion volume (0 and 7 mL g-1) showed amylose contents of 21.24 and 20.18%, respectively; a less compact endosperm, with individual starch granules interspaced with empty spaces; and pericarp thickness between 40.94 and 38.99 µm, respectively. By contrast, progenies with high popping expansion volume (30 and 35 mL g-1) showed amylose contents of 23.92 and 26.10%; a vitreous endosperm; more-compact starch granules without empty spaces in between; and pericarp thickness between 107.66 and 107.84 µm. Progenies with higher popping expansion volume exhibited a thicker pericarp, a high amylose percentage and a more-compact endosperm, whereas those with the lower expansion volumes showed a thinner pericarp, a lower amylose percentage and individual starch granules.(AU)

O milho-pipoca (Zea mays var. everta) tem um valor comercial mais elevado do que o milho comum e é um alimento popular entre os consumidores. Hoje, existe uma constante busca por cultivares com desempenho superior para os diversos caracteres de interesse, no caso da pipoca, produtividade e capacidade de expansão. O trabalho propõe caracterizar progênies de milho-pipoca com diferentes valores de capacidade de expansão quanto à composição química e micromorfologia. Foram estudados grãos do quinto ciclo (C5) de seleção recorrente intrapopulacional. As progênies foram selecionadas com base nos valores de capacidade de expansão de seus grãos. Os grãos foram quantificados para amilose e analisados quanto ao arranjo dos grânulos de amido e espessura do pericarpo por microscopia eletrônica de varredura. Progênies com baixa capacidade de expansão (0 e 7 mL g-1) apresentaram teor de amilose de 21,24 e 20,18% respectivamente; endosperma menos compacto, com grânulos de amido individualizados e intercalados com espaços vazios; e espessura do pericarpo medindo entre 40,94 e 38,99 µm, respectivamente. Por outro lado, progênies com alta capacidade de expansão (30 e 35 mL g-1) apresentaram teor de amilose de 23,92 e 26,10%, endosperma vítreo, sendo os grânulos de amido mais compactos e sem espaços vazios entre eles; e espessura do pericarpo medindo entre 107,66 e 107,84 µm. Progênies com altos valores de capacidade de expansão apresentam maior espessura de pericarpo, alta porcentagem de amilose e endosperma mais compactado, enquanto as com menores valores de expansão, apresentam menor espessura do pericarpo, menor porcentagem de amilose e grânulos de amido individualizados.(AU)

Genetic Screens to Target Embryo and Endosperm Pathways in Arabidopsis and Maize.

The major tissue types and stem-cell niches of plants are established during embryogenesis, and thus knowledge of embryo development is essential for a full understanding of plant development. Studies of seed development are also important for human health, because the nutrients stored in both the embryo and endosperm of plant seeds provide an essential part of our diet. Arabidopsis and maize have evolved different types of seeds, opening a range of experimental opportunities. Development of the Arabidopsis embryo follows an almost invariant pattern, while cell division patterns of maize embryos are variable. Embryo-endosperm interactions are also different between the two species: in Arabidopsis, the endosperm is consumed during seed development, while mature maize seeds contain an enormous endosperm. Genetic screens have provided important insights into seed development in both species. In the genomic era, genetic analysis will continue to provide important tools for understanding embryo and endosperm biology in plants, because single gene functional studies can now be integrated with genome-wide information. Here, we lay out important factors to consider when designing genetic screens to identify new genes or to probe known pathways in seed development. We then highlight the technical details of two previous genetic screens that may serve as useful examples for future experiments.

Characterization of the nucellus-specific dehydrin MdoDHN11 demonstrates its involvement in the tolerance to water deficit.

KEY MESSAGE: MdoDHN11 acts in the nucellus layer to protect the embryo and the endosperm from limited water availability during apple seed development. Dehydrins (DHNs) are protective proteins related to several plant developmental responses that involve dehydration such as seed desiccation and abiotic stresses. In apple (Malus × domestica Borkh.), the seed-specific MdoDHN11 was suggested to play important roles against dehydration during seed development. However, this hypothesis has not yet been evaluated. Within this context, several experiments were performed to functionally characterize MdoDHN11. In situ hybridization analysis during apple seed development showed that MdoDHN11 expression is confined to a maternal tissue called nucellus, a central mass of parenchyma between the endosperm and the testa. The MdoDHN11 protein was localized in the cytosol and nucleus. Finally, transgenic Arabidopsis plants expressing MdoDHN11 were generated and exposed to a severe water-deficit stress, aiming to mimic a situation that can occurs during seed development. All transgenic lines showed increased tolerance to water deficit in relation to wild-type plants. Taken together, our results provide evidences that MdoDHN11 plays important roles during apple seed development by protecting the embryo and the endosperm from limited water availability, and the mechanism of action probably involves the interaction of MdoDHN11 with proteins and other components in the cell.

Cytosolic calcium localization and dynamics during early endosperm development in the genus Agave (Asparagales, Asparagaceae).

Calcium is a secondary messenger that regulates and coordinates the cellular responses to environmental cues. Despite calcium being a key player during fertilization in plants, little is known about its role during the development of the endosperm. For this reason, the distribution, abundance, and dynamics of cytosolic calcium during the first stages of endosperm development of Agave tequilana and Agave salmiana were analyzed. Cytosolic calcium and actin filaments detected in the embryo sacs of Agave tequilana and A. salmiana revealed that they play an important role during the division and nuclear migration of the endosperm. After fertilization, a relatively high concentration of cytosolic calcium was located in the primary nucleus of the endosperm, as well as around migrating nuclei during the development of the endosperm. Cytosolic calcium participates actively during the first mitosis of the endosperm mother cell and interacts with the actin filaments that generate the motor forces during the migration of the nuclei through the large cytoplasm of the central cell.

Parboiling of sorghum grains as a strategy to improve endosperm yield and mineral content of refined flours.

The aims were (i) to optimize the parboiling/decorticating process by assessing the effects of parboiling soaking temperature (60-80 ℃) and pearling time (4-6 min and 5.5-9.5 min for red and white sorghum) on endosperm yield and ash content, to obtain pregelatinized refined flours from parboiling sorghum (RF-PS) and (ii) to evaluate the final content of selected nutrients in RF-PS obtained in these optimal conditions. Endosperm yield decreased with the increase of pearling time, and a maximum around 70 ℃ soaking temperature was observed. Ash content decreased with increasing soaking temperature and pearling time. The optimal conditions of soaking temperature-pearling time were 73.3 ℃-4.8 min for red sorghum and 67.9 ℃-8.6 min for white sorghum, considering maximum endosperm yield and ash content lower than 0.65 g/100 g. In RF-PS obtained under optimal conditions, protein, lipid, Cu, and free polyphenols were higher in red sorghum than white sorghum (10.16 ± 0.62 vs. 9.42 ± 0.61 g/100 g, 0.89 ± 0.16 vs. 0.62 ± 0.11 g/100 g, 2.08 ± 0.21 vs. 1.88 ± 0.20 mg/kg, 79.51 ± 14.51 vs. 63.82 ± 4.33 mg/100 g). Fe, Zn, Ca, and Na were higher in white sorghum than red sorghum (20.61 ± 2.92 vs. 17.56 ± 0.98 mg/kg, 11.94 ± 0.84 vs. 9.58 ± 1.65 mg/kg, 87.45 ± 12.91 vs. 75.31 ± 12.57 mg/kg, 129.62 ± 9.03 vs. 102.69 ± 28.34 mg/kg). No difference was observed in the contents of ash, Mg, and K. It was possible to improve endosperm yield using a parboiling process prior to decortication. In addition, this process contributes to increase the mineral content in RF-PS.

A new, harmless, high-throughput endosperm-based DNA extraction method for wheat / Um método de extração de DNA novo, inofensivo e de alto teor de endosperma para o trigo

In this study, a non-destructive, high-throughput, endosperm-based DNA extraction method was developed. To verify the non-destructive nature of this method, a germination test was performed on 288 seeds after sampling their endosperm, which gave a seedling emergence rate that was higher (97.6%) than that of the control group (92%). To confirm the feasibility of the new method, DNA was extracted from plants of a BC1F2 population by two different methods, namely, from endosperm using our rapid, high-throughput method (ER-DNA) and from young leaves emerging from the same sampled seed using the CTAB method (LC-DNA). The ER-DNA was undetectable by agarose gel electrophoresis, but was found to be an adequate replacement for LC-DNA for the amplification and detection of simple sequence repeats (SSRs). Further analysis revealed that ER-DNA was generally suitable for the generation of specific 500-750-bp fragments, but not for the amplification of 1,000-2,000-bp fragments. Our rapid, high-throughput method therefore has no deleterious effects on wheat seeds and yields DNA for SSR genotyping that is a suitable alternative to traditionally obtained DNA.(AU)

Neste estudo, foi desenvolvido um método de extração de DNA não destrutivo, de alto débito e endosperma. Para verificar a natureza não destrutiva deste método, um teste de germinação foi realizado em 288 sementes após a amostragem do endosperma, o que deu uma taxa de emergência de plântulas maior (97,6%) do que a do grupo controle (92%). Para confirmar a viabilidade do novo método, o DNA foi extraído de plantas de uma população de BC1F2 por dois métodos diferentes, a saber, do endosperma usando nosso método rápido de alto rendimento (ER-DNA) e de folhas jovens que emergem da mesma semente amostrada usando o método CTAB (LC-DNA). O ER-DNA foi indetectável por eletroforese em gel de agarose, mas foi encontrado como uma substituição adequada para LC-DNA para a amplificação e detecção de repetições simples de seqüência (SSRs). Uma análise posterior revelou que o ER-DNA era geralmente adequado para a geração de fragmentos específicos de 500-750 pb, mas não para a amplificação de fragmentos de 1.000-2.000 pb. Nosso método rápido e de alto débito, portanto, não tem efeitos deletérios sobre as sementes de trigo e produz DNA para a genotipagem de SSR que é uma alternativa adequada ao DNA obtido tradicionalmente.(AU)