Cell wall remodeling under salt stress: Insights into changes in polysaccharides, feruloylation, lignification, and phenolic metabolism in maize.

Although cell wall polymers play important roles in the tolerance of plants to abiotic stress, the effects of salinity on cell wall composition and metabolism in grasses remain largely unexplored. Here, we conducted an in-depth study of changes in cell wall composition and phenolic metabolism induced upon salinity in maize seedlings and plants. Cell wall characterization revealed that salt stress modulated the deposition of cellulose, matrix polysaccharides and lignin in seedling roots, plant roots and stems. The extraction and analysis of arabinoxylans by size-exclusion chromatography, 2D-NMR spectroscopy and carbohydrate gel electrophoresis showed a reduction of arabinoxylan content in salt-stressed roots. Saponification and mild acid hydrolysis revealed that salinity also reduced the feruloylation of arabinoxylans in roots of seedlings and plants. Determination of lignin content and composition by nitrobenzene oxidation and 2D-NMR confirmed the increased incorporation of syringyl units in lignin of maize roots. Salt stress also induced the expression of genes and the activity of enzymes enrolled in phenylpropanoid biosynthesis. The UHPLC-MS-based metabolite profiling confirmed the modulation of phenolic profiling by salinity and the accumulation of ferulate and its derivatives 3- and 4-O-feruloyl quinate. In conclusion, we present a model for explaining cell wall remodeling in response to salinity.

Leaf application of chitosan and physiological evaluation of maize hybrids contrasting for drought tolerance under water restriction / Aplicação foliar de quitosana e avaliação fisiológica de híbridos de milho contrastantes para tolerância à seca sob restrição hídrica

It is a fact that the regions that cultivate the most maize crop do not have fully adequate technologies to measure productivity losses caused by irregularities in water availability. The objective of this study was to evaluate the physiological characteristics of maize hybrids tolerant (DKB 390) and sensitive (BRS 1030) to drought, at V5 growth stage and under water restriction, in order to understand the mechanisms involved in the induction of tolerance to drought by chitosan in contrasting maize genotypes. Plants were cultivated in pots at a greenhouse, and chitosan 100 ppm was applied by leaf spraying. The water restriction was imposed for 10 days and then leaf gaseous exchange and chlorophyll fluorescence were evaluated. The tolerant hybrid (DKB 390) showed higher photosynthesis, stomatal conductance, carboxylation efficiency, electron transport rate, and non-photochemical quenching when chitosan was used. Plants from tolerant genotype treated with chitosan were more tolerant to water stress because there were more responsive to the biopolymer.(AU)

As regiões que cultivam milho como cultura principal ainda não possuem tecnologias adequadas para mensurar as perdas na produtividade decorrentes na disponibilidade irregular de água. O objetivo desse estudo foi avaliar as características fisiológicas de híbridos de milho tolerante (DKB 390) e sensível (BRS1030) à seca, no estádio de crescimento V5 e sob restrição hídrica, para compreender os mecanismos envolvidos na indução de tolerância à seca pela quitosana em genótipos contrastantes. As plantas foram cultivadas vasos na casa de vegetação e a quitosana 100 ppm foi aplicada por pulverização foliar. A restrição hídrica durou 10 dias e foram avaliadas as trocas gasosas e a fluorescência da clorofila. O híbrido tolerante (DKB 390) apresentou maior fotossíntese, condutância estomática, eficiência de carboxilação, taxa de transporte de elétrons e quenching não fotoquímico quando aplicada a quitosana. As plantas do genótipo tolerante tratadas com quitosana foram mais tolerantes ao déficit hídrico porque foram mais responsivas ao biopolímero.(AU)

Apparent nutrient digestibility of carbohydrate (corn) in Cirrhinus mrigala under the influence of chromium chloride hexahydrate / Digestibilidade aparente de nutrientes oriundos de carboidratos (milho) em Cirrhinus mrigala sob a influência do cloreto de cromo hexa-hidratado

A feeding trial was carried out to assess the effect of dietary chromium supplementation on apparent nutrient digestibility coefficient (%) of gelatinized and non-gelatinized corn in Cirrhinus mrigala fingerlings for 90 days. Using various levels of chromium chloride hexahydrate six test diets designated as T1 (G/0.0 Cr2Cl3.6H2O mg Kg-1), T2 (NG/0.0 Cr2Cl3.6H2O mg Kg-1), T3 (G/0.2 Cr2Cl3.6H2O mg Kg-1), T4 (NG/0.2 Cr2Cl3.6H2O mg Kg-1), T5 (G/0.4 Cr2Cl3.6H2O mg Kg-1) and T6 (NG/0.4 Cr2Cl3.6H2O mg Kg-1) were prepared. Results showed highest apparent digestibility coefficient (ADC) of nutrients dry matter, crude lipid and gross energy in test diet T5 that was gelatinized and supplemented with chromium 0.4 mg/Kg while, for crude protein higher value of nutrient digestibility was recorded in T3 test diet (G/0.2 Cr2Cl3.6H2O mg/Kg). It was concluded that chromium supplementation with gelatinized corn in fish (Cirrhinus mrigala) diet can improve the nutrients digestibility more efficiently.(AU)

Um experimento referente à alimentação foi realizado para avaliar o efeito da suplementação dietética de cromo, no coeficiente de digestibilidade aparente do nutriente (%) do milho gelatinizado e não gelatinizado em alevinos de Cirrhinus mrigala por 90 dias. Usando vários níveis de cloreto de cromo hexa-hidratado, seis dietas testes foram designadas e preparadas como: T1 (G/0,0 Cr2Cl3.6H2O mg Kg-1); T2 (NG/0,0 Cr2Cl3.6H2O mg Kg-1); T3 (G/0,2 Cr2Cl3.6H2O mg Kg-1); T4 (NG/0,2 Cr2Cl3.6H2O mg Kg-1); T5 (G/0,4 Cr2Cl3.6H2O mg Kg-1) e T6 (NG/0,4 Cr2Cl3.6H2O mg Kg-1). Os resultados mostraram maior coeficiente de digestibilidade aparente (CDA) em matéria seca de nutrientes, lipídios brutos e energia bruta na dieta teste T5, que foi gelatinizada e suplementada com cromo 0,4 mg / Kg; enquanto que para a proteína bruta, foi registrada maior digestibilidade dos nutrientes na dieta teste T3 (G/0,2 Cr2Cl3.6H2O mg/Kg). Concluiu-se que a suplementação de cromo com milho gelatinizado na dieta de peixes (Cirrhinus mrigala) pôde melhorar a digestibilidade dos nutrientes de forma mais eficiente.(AU)

Evaluation of the effects of humic acids on maize root architecture by label-free proteomics analysis.

Humic substances have been widely used as plant growth promoters to improve the yield of agricultural crops. However, the mechanisms underlying this effect remain unclear. Root soluble protein profiles in plants 11 days after planting and cultivated with and without humic acids (HA, 50 mg CL-1), were analyzed using the label-free quantitative proteomic approach. Cultivation of maize with HA resulted in higher fresh weight of roots than in untreated plants (control). Plants treated with HA showed increased number, diameter and length of roots. In the proteomics analysis, differences were detected in the following categories: energy metabolism, cytoskeleton, cellular transport, conformation and degradation of proteins, and DNA replication. Thirty-four proteins were significantly more abundant in the seedlings treated with HA, whereas only nine proteins were abundant in the control. The effects on root architecture, such as the induction of lateral roots and biomass increase were accompanied by changes in the energy metabolism-associated proteins. The results show that the main effect of HA is protective, mainly associated with increased expression of the 2-cys peroxidase, putative VHS/GAT, and glutathione proteins. Indeed, these proteins had the highest fold-difference. Overall, these results improve our understanding of the molecular mechanisms of HA-promoted plant growth.

Lanthanum content and effects on growth, gas exchanges, and chlorophyll index in maize plants / Teor de lantânio e efeitos no crescimento, trocas gasosas e teor de clorofila em plantas de milho

Rare earth elements, such as lanthanum (La), have been applied to agriculture via fertilizers, aiming to increase the productivity and crop quality, such results observed mainly in China. However, the knowledge about the effect of La on maize growth, as well as for other species, despite the growing interest it is still limited. The aim of this study was to evaluate the effects of La on maize growth, La content,photosynthetic rate and chlorophyll content in maize plants in response to La treatment (0, 25, 50, 100, 150, 300 and 600 μM) in nutrient solution for three weeks. The plants were placed in geminated pots using a split-root technique. One of the pots in the geminated set was filled with a complete nutrient solution without La, while another was filled with a nutrient solution without phosphorus but containing different concentrations of La. It was verified that roots of maize plants can accumulate approximately sixty percent more La than shoots. Moreover, low La concentrations stimulated an increase in chlorophyll index,resulting in a slight increase in shoot biomass. At higher levels, La didnt reduce growth but caused adecrease in both photosynthetic rate and chlorophyll index.(AU)

Elementos terra rara, como lantânio (La), são aplicados à agricultura via fertilizantes, com oobjetivo de aumentar a produtividade e a qualidade das culturas agrícolas, fato observado principalmente naChina. Entretanto, o conhecimento sobre o efeito de La no crescimento do milho, bem como para outrasculturas agrícolas, ainda é limitado apesar do interesse crescente. O presente estudo objetivou avaliar oefeito da aplicação de La sobre o crescimento, teor de La, fotossíntese e teor de clorofila em plantas demilho, expostas a concentrações crescentes de La (0, 25, 50, 100, 150, 300 e 600 μM) em solução nutritivapor três semanas. As plantas foram colocadas em vasos geminados usando uma técnica de raiz divisória.Um dos potes no conjunto geminado foi preenchido com solução nutritiva completa que não continha La,enquanto outra foi preenchida com solução nutritiva sem fósforo, mas contendo diferentes concentraçõesde La. Foi verificado que as raízes das plantas de milho podem acumular sessenta por cento mais La quandocomparado à parte aérea. Além disso, as baixas concentrações de La estimularam o aumento do teor declorofila, resultando em um aumento na biomassa da parte aérea. Em concentrações mais elevadas, aaplicação de La não reduziu o crescimento das plantas de milho, mas causou diminuição na taxafotossintética e no índice de clorofila.(AU)

Alterations in Heterochromatic Knobs in Maize Callus Culture by Breakage-Fusion-Bridge Cycle and Unequal Crossing Over.

The meiotic and mitotic behavior of regenerated plants derived from a long-term callus culture, designated 12-F, was analyzed. This culture was heterozygous for an amplification of the heterochromatic knob on the long arm of chromosome 7 (K7L). We aimed to investigate if the amplification resulted from a breakage-fusion-bridge (BFB) cycle or from unequal sister chromatid recombination. Therefore, C-banded mitotic metaphases and pachytene, diakinesis, and anaphase I of regenerated plants were analyzed. Additionally, the occurrence of alterations in K7L was investigated in C-banded metaphases from short-term callus cultures derived from lines related to the donor genotype of the 12-F culture. As a result, plants homozygous and heterozygous for the amplification were detected. Meiosis was normal with few abnormalities, such as a low frequency of univalents at diakinesis. In the callus cultures a chromosome 7 with knobs of different sizes in the sister chromatids was detected and interpreted as a result of unequal crossing over. Other chromosomal alterations were consistent with the occurrence of BFB cycles. The finding of unequal crossing over in the cultures supports the conclusion that the amplification in the culture 12-F would be derived from this mechanism. If the amplification was derived from a BFB cycle, the terminal euchromatic segment between knob and the telomere would be deleted, and possibly, homozygous plants would not be viable.

Lanthanum content and effects on growth, gas exchanges, and chlorophyll index in maize plants / Teor de lantânio e efeitos no crescimento, trocas gasosas e teor de clorofila em plantas de milho

Rare earth elements, such as lanthanum (La), have been applied to agriculture via fertilizers, aiming to increase the productivity and crop quality, such results observed mainly in China. However, the knowledge about the effect of La on maize growth, as well as for other species, despite the growing interest it is still limited. The aim of this study was to evaluate the effects of La on maize growth, La content,photosynthetic rate and chlorophyll content in maize plants in response to La treatment (0, 25, 50, 100, 150, 300 and 600 μM) in nutrient solution for three weeks. The plants were placed in geminated pots using a split-root technique. One of the pots in the geminated set was filled with a complete nutrient solution without La, while another was filled with a nutrient solution without phosphorus but containing different concentrations of La. It was verified that roots of maize plants can accumulate approximately sixty percent more La than shoots. Moreover, low La concentrations stimulated an increase in chlorophyll index,resulting in a slight increase in shoot biomass. At higher levels, La didn’t reduce growth but caused adecrease in both photosynthetic rate and chlorophyll index.

Elementos terra rara, como lantânio (La), são aplicados à agricultura via fertilizantes, com oobjetivo de aumentar a produtividade e a qualidade das culturas agrícolas, fato observado principalmente naChina. Entretanto, o conhecimento sobre o efeito de La no crescimento do milho, bem como para outrasculturas agrícolas, ainda é limitado apesar do interesse crescente. O presente estudo objetivou avaliar oefeito da aplicação de La sobre o crescimento, teor de La, fotossíntese e teor de clorofila em plantas demilho, expostas a concentrações crescentes de La (0, 25, 50, 100, 150, 300 e 600 μM) em solução nutritivapor três semanas. As plantas foram colocadas em vasos geminados usando uma técnica de raiz divisória.Um dos potes no conjunto geminado foi preenchido com solução nutritiva completa que não continha La,enquanto outra foi preenchida com solução nutritiva sem fósforo, mas contendo diferentes concentraçõesde La. Foi verificado que as raízes das plantas de milho podem acumular sessenta por cento mais La quandocomparado à parte aérea. Além disso, as baixas concentrações de La estimularam o aumento do teor declorofila, resultando em um aumento na biomassa da parte aérea. Em concentrações mais elevadas, aaplicação de La não reduziu o crescimento das plantas de milho, mas causou diminuição na taxafotossintética e no índice de clorofila.

Small RNA differential expression and regulation in Tuxpeño maize embryogenic callus induction and establishment.

Somatic embryogenesis represents an alternative developmental process used to achieve genetic transformation and to approach key questions in maize development. It is known that embryogenic callus induction and plant regeneration are accompanied by microRNA expression changes. However, small RNA (sRNA) populations have not been explored during the proliferative callus subculture establishment and their impact on maintaining the dedifferentiated status and embryogenic potential is far from being completely understood. Here we globally tested the sRNA populations in explants (immature embryos), induced and established maize embryogenic callus from the Mexican cultivar VS-535, Tuxpeño landrace. We detected readjustments in 24 nt and 21-22 nt sRNAs during the embryogenic callus (EC) establishment and maintenance. A follow up on specific microRNAs (miRNAs) indicated that miRNAs related to stress response substantially increase upon the callus proliferation establishment, correlating with a reduction in some of their target levels. On the other hand, while 24 nt-long heterochromatic small interfering RNAs (hc-siRNAs) derived from transposable retroelements transiently decreased in abundance during the EC establishment, a population of 22 nt-hc-siRNAs increased. This was accompanied by reduction in transposon expression in the established callus subcultures. We conclude that stress- and development-related miRNAs are highly expressed upon maize EC callus induction and during maintenance of the subcultures, while miRNAs involved in hormone response only transiently increase during induction. In addition, the establishment of a proliferative status in embryogenic callus is accompanied by important readjustments in hc-siRNAs mapping to long tandem repeat (LTR) retrotransposons, and their expression regulation.

Economic analysis of corn inoculated with Azospirillum brasilense associated with nitrogen sources and doses / Análise econômica do milho inoculado com Azospirillum brasilense associado a doses e fontes de nitrogênio

Azospirillum brasilense is a bacterium known for its biological nitrogen fixation (BNF) in corn crops. However, there is a lack of comprehensive research defining how much mineral N should be applied to maximize the efficiency of BNF and attain high, economically sustainable yields. Moreover, it would be interesting to investigate whether adding urea with NBPT urease inhibitor might increase BNF in grasses. Therefore, the aim of this study was to determine the effect of inoculation with Azospirillum brasilense associated with N sources and doses in a Cerrado biome soil by evaluating the grain yield of irrigated corn in economic terms. The experiment was conducted in Selvíria, MS, Brazil under a no-till system on a Latossolo Vermelho distrófico (Oxisol). The experiment was set up as a randomized block design with four replications in a 2 × 5 × 2 factorial arrangement consisting of two sources of N (urea and urea with NBPT urease enzyme inhibitor) and five N doses applied as top-dressing (0, 50, 100, 150, and 200 kg ha-1), with and without the inoculation of seeds with A. brasilense. Inoculation with Azospirillum brasilense makes corn growth much more profitable, irrespective of the dose and source of N. Addition of 200 kg ha-1 N in the form of conventional urea coupled with inoculation with Azospirillum brasilense increases grain yield; however, the highest economic return is obtained with Napplied at 100 kg ha-1 with conventional urea and inoculation.(AU)

O Azospirillum brasilense se destaca na fixação biológica de nitrogênio (FBN) em cultivo de milho. Contudo, faltam maiores pesquisas para definição do quanto se pode aplicar de N mineral quanto à eficiência da FBN, para alcançar altas produtividades economicamente sustentáveis. Além disso, seria interessante analisar se a ureia com inibidor da enzima urease NBPT pode ser menos prejudicial, beneficiando a FBN em gramíneas. Sendo assim, objetivou-se estudar o efeito da inoculação com Azospirillum brasilense, associado às doses e fontes de N, em solo de Cerrado, avaliando economicamente a produtividade de grãos de milho irrigado. O experimento foi desenvolvido em Selvíria - MS, em sistema plantio direto, em Latossolo Vermelho distrófico. O delineamento experimental foi em blocos casualizados, com quatro repetições, dispostos em um esquema fatorial 2 x 5 x 2, sendo: duas fontes de N (ureia e ureia com inibidor da enzima urease NBPT); cinco doses de N em cobertura (0, 50, 100, 150 e 200 kg ha-1); com e sem inoculação das sementes com A. brasilense. A inoculação com Azospirillum brasilense torna o cultivo de milho mais lucrativo, independentemente da dose e fonte de N. A dose de 200 kg ha-1 de N na forma de ureia convencional associado à inoculação com Azospirillum brasilense proporciona maior produtividade de grãos, entretanto, o maior retorno econômico é obtido na dose de 100 kg ha-1 de N, com ureia convencional e inoculação.(AU)

Economic analysis of corn inoculated with Azospirillum brasilense associated with nitrogen sources and doses / Análise econômica do milho inoculado com Azospirillum brasilense associado a doses e fontes de nitrogênio

Azospirillum brasilense is a bacterium known for its biological nitrogen fixation (BNF) in corn crops. However, there is a lack of comprehensive research defining how much mineral N should be applied to maximize the efficiency of BNF and attain high, economically sustainable yields. Moreover, it would be interesting to investigate whether adding urea with NBPT urease inhibitor might increase BNF in grasses. Therefore, the aim of this study was to determine the effect of inoculation with Azospirillum brasilense associated with N sources and doses in a Cerrado biome soil by evaluating the grain yield of irrigated corn in economic terms. The experiment was conducted in Selvíria, MS, Brazil under a no-till system on a Latossolo Vermelho distrófico (Oxisol). The experiment was set up as a randomized block design with four replications in a 2 × 5 × 2 factorial arrangement consisting of two sources of N (urea and urea with NBPT urease enzyme inhibitor) and five N doses applied as top-dressing (0, 50, 100, 150, and 200 kg ha-1), with and without the inoculation of seeds with A. brasilense. Inoculation with Azospirillum brasilense makes corn growth much more profitable, irrespective of the dose and source of N. Addition of 200 kg ha-1 N in the form of conventional urea coupled with inoculation with Azospirillum brasilense increases grain yield; however, the highest economic return is obtained with Napplied at 100 kg ha-1 with conventional urea and inoculation.

O Azospirillum brasilense se destaca na fixação biológica de nitrogênio (FBN) em cultivo de milho. Contudo, faltam maiores pesquisas para definição do quanto se pode aplicar de N mineral quanto à eficiência da FBN, para alcançar altas produtividades economicamente sustentáveis. Além disso, seria interessante analisar se a ureia com inibidor da enzima urease NBPT pode ser menos prejudicial, beneficiando a FBN em gramíneas. Sendo assim, objetivou-se estudar o efeito da inoculação com Azospirillum brasilense, associado às doses e fontes de N, em solo de Cerrado, avaliando economicamente a produtividade de grãos de milho irrigado. O experimento foi desenvolvido em Selvíria - MS, em sistema plantio direto, em Latossolo Vermelho distrófico. O delineamento experimental foi em blocos casualizados, com quatro repetições, dispostos em um esquema fatorial 2 x 5 x 2, sendo: duas fontes de N (ureia e ureia com inibidor da enzima urease NBPT); cinco doses de N em cobertura (0, 50, 100, 150 e 200 kg ha-1); com e sem inoculação das sementes com A. brasilense. A inoculação com Azospirillum brasilense torna o cultivo de milho mais lucrativo, independentemente da dose e fonte de N. A dose de 200 kg ha-1 de N na forma de ureia convencional associado à inoculação com Azospirillum brasilense proporciona maior produtividade de grãos, entretanto, o maior retorno econômico é obtido na dose de 100 kg ha-1 de N, com ureia convencional e inoculação.

ZmMBD101 is a DNA-binding protein that maintains Mutator elements chromatin in a repressive state in maize.

In maize (Zea mays), as well as in other crops, transposable elements (TEs) constitute a great proportion of the genome. Chromatin modifications play a vital role in establishing transposon silencing and perpetuating the acquired repressive state. Nucleosomes associated with TEs are enriched for dimethylation of histone H3 at lysine 9 and 27 (H3K9me2 and H3K27me2, respectively), signals of repressive chromatin. Here, we describe a chromatin protein, ZmMBD101, involved in the regulation of Mutator (Mu) genes in maize. ZmMBD101 is localized to the nucleus and contains a methyl-CpG-binding domain (MBD) and a zinc finger CW (CW) domain. Transgenic lines with reduced levels of ZmMBD101 transcript present enhanced induction of Mu genes when plants are irradiated with UV-B. Chromatin immunoprecipitation analysis with H3K9me2 and H3K27me2 antibodies indicated that ZmMBD101 is required to maintain the levels of these histone repressive marks at Mu terminal inverted repeats (TIRs) under UV-B conditions. Although Mutator inactivity is associated with DNA methylation, cytosine methylation at Mu TIRs is not affected in ZmMBD101 deficient plants. Several plant proteins are predicted to share the simple CW-MBD domain architecture present in ZmMBD101. We hypothesize that plant CW-MBD proteins may also function to protect plant genomes from deleterious transposition.

A bioassay for brassinosteroid activity based on the in vitro fluorimetric detection of nitric oxide production.

Recent studies have shown that low concentrations of brassinolide induce a rapid generation of nitric oxide in mesophyll cells of maize leaves, which can be easily detected by fluorimetric methods. In this work we describe a series of natural and synthetic brassinosteroids that are able to trigger in vitro NO production in tomato cells that exhibits dose-response behavior. We propose that this effect can be used to develop a new rapid and very sensitive bioassay for brassinosteroid activity that offers several advantages when compared to the current methodologies.

Continuous, high-resolution biospeckle imaging reveals a discrete zone of activity at the root apex that responds to contact with obstacles.

BACKGROUND AND AIMS: Shining a laser onto biological material produces light speckles termed biospeckles. Patterns of biospeckle activity reflect changes in cell biochemistry, developmental processes and responses to the environment. The aim of this work was to develop methods to investigate the biospeckle activity in roots and to characterize the distribution of its intensity and response to thigmostimuli. METHODS: Biospeckle activity in roots of Zea mays, and also Jatropha curcas and Citrus limonia, was imaged live and in situ using a portable laser and a digital microscope with a spatial resolution of 10 µm per pixel and the ability to capture images every 0.080 s. A procedure incorporating a Fujii algorithm, image restoration using median and Gaussian filters, image segmentation using maximum-entropy threshold methods and the extraction of features using a tracing algorithm followed by spline fitting were developed to obtain quantitative information from images of biospeckle activity. A wavelet transform algorithm was used for spectral decomposition of biospeckle activity and generalized additive models were used to attribute statistical significance to changes in patterns of biospeckle activity. KEY RESULTS: The intensity of biospeckle activity was greatest close to the root apex. Higher frequencies (3-6 Hz) contributed most to the total intensity of biospeckle activity. When a root encountered an obstacle, the intensity of biospeckle activity decreased abruptly throughout the root system. The response became attenuated with repeated thigmostimuli. CONCLUSIONS: The data suggest that at least one component of root biospeckle activity resulted from a biological process, which is located in the zone of cell division and responds to thigmostimuli. However, neither individual cell division events nor root elongation is likely to be responsible for the patterns of biospeckle activity.

Insights into the TOR-S6K signaling pathway in maize (Zea mays L.). pathway activation by effector-receptor interaction.

The primordial TOR pathway, known to control growth and cell proliferation, has still not been fully described for plants. Nevertheless, in maize, an insulin-like growth factor (ZmIGF) peptide has been reported to stimulate this pathway. This research provides further insight into the TOR pathway in maize, using a biochemical approach in cultures of fast-growing (FG) and slow-growing (SG) calli, as a model system. Our results revealed that addition of either ZmIGF or insulin to SG calli stimulated DNA synthesis and increased the growth rate through cell proliferation and increased the rate of ribosomal protein (RP) synthesis by the selective mobilization of RP mRNAs into polysomes. Furthermore, analysis of the phosphorylation status of the main TOR and S6K kinases from the TOR pathway revealed stimulation by ZmIGF or insulin, whereas rapamycin inhibited its activation. Remarkably, a putative maize insulin-like receptor was recognized by a human insulin receptor antibody, as demonstrated by immunoprecipitation from membrane protein extracts of maize callus. Furthermore, competition experiments between ZmIGF and insulin for the receptor site on maize protoplasts suggested structural recognition of the putative receptor by either effector. These data were confirmed by confocal immunolocalization within the cell membrane of callus cells. Taken together, these data indicate that cell growth and cell proliferation in maize depend on the activation of the TOR-S6K pathway through the interaction of an insulin-like growth factor and its receptor. This evidence suggests that higher plants as well as metazoans have conserved this biochemical pathway to regulate their growth, supporting the conclusion that it is a highly evolved conserved pathway.

Metamorphosis of the Basidiomycota Ustilago maydis: transformation of yeast-like cells into basidiocarps.

Ustilago maydis (DC) Cda., a phytopathogenic Basidiomycota, is the causal agent of corn smut. During its life cycle U. maydis alternates between a yeast-like, haploid nonpathogenic stage, and a filamentous, dikaryotic pathogenic form that invades the plant and induces tumor formation. As all the members of the Subphylum Ustilaginomycotina, U. maydis is unable to form basidiocarps, instead it produces teliospores within the tumors that germinate forming a septate basidium (phragmobasidium). We have now established conditions allowing a completely different developmental program of U. maydis when grown on solid medium containing auxins in dual cultures with maize embryogenic calli. Under these conditions U. maydis forms large hemi-spheroidal structures with all the morphological and structural characteristics of gastroid-type basidiocarps. These basidiocarps are made of three distinct hyphal layers, the most internal of which (hymenium) contains non-septate basidia (holobasidia) from which four basidiospores develop. In basidiocarps meiosis and genetic recombination occur, and meiotic products (basidiospores) segregate in a Mendelian fashion. These results are evidence of sexual cycle completion of an Ustilaginomycotina in vitro, and the demonstration that, besides its quasi-obligate biotrophic pathogenic mode of life, U. maydis possesses the genetic program to form basidiocarps as occurs in saprophytic Basidiomycota species.

ms17: a meiotic mutation causing partial male sterility in a corn silage hybrid.

Cytological analysis under light microscopy of the single hybrid P30R50 of silage corn revealed an abnormal pattern of microsporogenesis that affected the meiotic products. Meiosis progressed normally until diakinesis, but before migration to the metaphase plate, bivalents underwent total desynapsis and 20 univalent chromosomes were scattered in the cytoplasm. At this stage, meiocytes also exhibited a number of chromatin-like fragments scattered throughout the cell. Metaphase I was completely abnormal in the affected cells, and univalent chromosomes and fragments were distributed among several curved spindles. Anaphase I did not occur, and each chromosome or group of chromosomes originated a micronucleus. After this phase, an irregular cytokinesis occurred, and secondary meiocytes with several micronuclei were observed. Metaphase II and anaphase II also did not occur, and after the second cytokinesis, the genomes were fractionated into polyads, generating several unbalanced microspores, with various-sized nuclei. About 35% of the tetrads were abnormal in the hybrid. This spontaneous mutation had been previously reported in a USA maize line called ms17 and was found to cause male sterility.

Effects of low-energy argon ion implantation on the dynamic organization of the actin cytoskeleton during maize pollen germination.

The relationship between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination is a central theme in plant reproductive biology research. Maize (Zea mays) pollen grains were implanted with 30 keV argon ion (Ar(+)) beams at doses ranging from 0.78 x 10(15) to 13 x 10(15) ions/cm(2). The effects of low-energy ion implantation on pollen germination viability and the dynamic organization of the actin cytoskeleton during pollen germination were studied using confocal laser scanning microscopy. Maize pollen germination rate increased remarkably with Ar(+) dose, in the range from 3.9 x 10(15) to 6.5 x 10(15) ions/cm(2); the germination rate peaked at an Ar(+) dose of 5.2 x 10(15) ions/cm(2). When the implantation dose exceeded 7.8 x 10(15) ions/cm(2), the rate of pollen germination decreased sharply. The actin filaments assembled in pollen grains implanted with 5.2 x 10(15) ions/cm(2) Ar(+) much earlier than in controls. The actin filaments organized as longer parallel bundles and extended into the emerging pollen tube in treated pollen grains, while they formed random and loose fine bundles and were gathered at the pollen aperture in the control. The reorganization of actin cytoskeleton in the pollen implanted with 9.1 x 10(15) ions/cm(2) Ar(+) was slower than in controls. There was a positive correlation between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination. Ion implantation into pollen did not cause changes in the polarization of actin filaments and organelle dynamics in the pollen tubes. The effects of Ar(+) implantation on pollen germination could be mediated by changes in the polymerization and rearrangement of actin polymers.

Nitric oxide mediates humic acids-induced root development and plasma membrane H+-ATPase activation.

It is widely reported that some humic substances behave as exogenous auxins influencing root growth by mechanisms that are not yet completely understood. This study explores the hypothesis that the humic acids' effects on root development involve a nitric oxide signaling. Maize seedlings were treated with HA 20 mg C L(-1), IAA 0.1 nM, and NO donors (SNP or GSNO), in combination with either the auxin-signaling inhibitor PCIB, the auxin efflux inhibitor TIBA, or the NO scavenger PTIO. H(+)-transport-competent plasma membrane vesicles were isolated from roots to investigate a possible link between NO-induced H(+)-pump and HA bioactivity. Plants treated with either HA or SNP stimulated similarly the lateral roots emergence even in the presence of the auxin inhibitors, whereas NO scavenger diminished this effect. These treatments induced H(+)-ATPase stimulation by threefold, which was abolished by PTIO and decreased by auxin inhibitors. HA-induced NO synthesis was also detected in the sites of lateral roots emergence. These data depict a new scenario where the root development stimulation and the H(+)-ATPase activation elicited by either HA or exogenous IAA depend essentially on mechanisms that use NO as a messenger induced site-specifically in the early stages of lateral root development.

Differences in intracellular localization of corn stunt spiroplasmas in magnesium treated maize.

Maize plants infected with Spiroplasma kunkelii show symptoms similar to that of plants in a magnesium-deficient soil, and it has been shown that the spiroplasma alters the plants' magnesium absorption. In the current study we compared changes associated to either spiroplasma infection, two soil magnesium levels and their combinations. Plant symptoms were recorded and correlated with transmission electron microscopy observations. Plants grown on a high magnesium treatment showed no macroscopical alterations nor organelle ultrastructural alterations, while plants on a low magnesium treatment showed macroscopical vein yellowing and, ultrastructurally, they had most chloroplasts and mitochondrial membranes altered. Infected plants on a low magnesium treatment had an ageing aspect, ultrastructurally showed chloroplasts and mitochondrial alterations similar to those non-infected and grown on a low magnesium treatment, and spiroplasma cells were found in phloem cells, but outside their cytoplasm. Infected plants on a high magnesium treatment showed similar symptoms and ultrastructural alterations as either non-infected plants on the low magnesium treatment or in infected plants on the low magnesium treatment, but differ from them in that the spiroplasma cells were located inside the cytoplasm. Results suggest that magnesium is involved in the plant-pathogen interaction.

Distribution of linker histone variants during plant cell differentiation in the developmental zones of the maize root, dedifferentiation in callus culture after auxin treatment.

Although several linker histone variants have been studied in both animal and plant organisms, little is known about their distribution during processes that involve alterations in chromatin function, such as differentiation, dedifferentiation and hormone treatment. In this study, we identified linker histone variants by using specific anti-histone Hl antibodies. Each variant's ratio to total Hl in the three developmental zones of maize (Zea mays L.) root and in callus cultures derived from them was estimated in order to define possible alterations either during plant cell differentiation or during their dedifferentiation. We also evaluated linker histone variants' ratios in the developmental zones of maize roots treated with auxin in order to examine the effects of exogenous applied auxin to linker histone variant distribution. Finally, immunohistochemical detection was used to identify the root tissues containing each variant and correlate them with the physiological status of the plant cells. According to the results presented in this study, linker histone variants' ratios are altered in the developmental zones of maize root, while they are similar to the meristematic zone in samples from callus cultures and to the differentiation zone in samples from roots treated with auxin. We propose that the alterations in linker histone variants' ratios are correlated with plant cell differentiation and dedifferentiation.