Bio-detoxification of mycotoxins by lactic acid bacteria from different food matrices

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP- PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 g/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 m pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.

Creaciones fitogenéticas en Maíz con respuesta adaptativa a la zona marginal

La creación de una población mejorada de maíz, requiere al menos tres etapas: a) La conformación de una "población original" con variabilidad genética, b) La selección preliminar de plantas y/o progenies, y c) La "evaluación" en condiciones de campo de las mejores descendencias integradas en poblaciones experimentales. Precisamente, este proyecto está dirigido a la creación de variedades con estabilidad de rendimientos en las condiciones ambientales de la zona centro-norte de la Provincia de Córdoba, en donde el maíz es el único cereal que se cosecha y una de las mejores posibilidades como forrajera estival. Se ha inscripto el cv. "MPB-FCA 856", un maíz colorado duro creado para la producción de granos, que en marcado déficit hídrico produce rendimientos superiores a otros cultivares. Actualmente se dispone de una variedad sintética, próxima a ser inscripta como una nueva creación. Con orientación al desarrollo de otros cultivares, se plantean las líneas de trabajo que se enumeran seguidamente: Tema 1. Maíz Colorado Duro: Obtención de una variedad sintética. Se iniciará la caracterización varietal a fin de inscribir en el Instituto Nacional de Semillas el nuevo cv. sintético formado por 10 líneas S1, que presenta altos rendimientos de granos. Tema 2. Maíz Forrajero: Desarrollo de un cultivar con aptitud para ser utilizado como forraje en planta. En la zona marginal el pastoreo de lotes de maíz es común tanto en años con pocas precipitaciones pluviales como en ciclos favorables, por cuanto es una muy buena alternativa de forraje estival tanto en verdeo, silo o diferido. Se dispone de líneas propias con características de alto macollamiento y capacidad de rebrote; se plantea su mejoramiento por caracteres que hacen a la calidad forrajera, tales como porcentaje de materia seca, palatabilidad, fibra bruta, etc. Tema 3. Mejora en la eficiencia de selección para tolerancia a estrés hídrico en maíz ( Zea mays L.). Aportar evidencia experimental que mejore la eficiencia de la selección, cuando los genotipos se dispersarán en una gama de ambientes contrastantes por su disponibilidad hídrica. Se comenzará un esquema de selección recurrente en ambientes con diferente disponibilidad hídrica, empleando características que permitan identificar genotipos tolerantes. Paralelamente se implementarán metodologías de evaluación del germoplasma disponible mediante pruebas de laboratorio y campo, a fin de detectar fuentes de genes para tolerancia al estrés hídrico.

Mapeo de QTL para Mal de Río Cuarto en maíz. QTL mapping for tolerance to Mal de Río Cuarto disease in maize.

El Mal de Río Cuarto (MRC) es una enfermedad del maíz (Zea mays L.), endémica de ciertas zonas de la Argentina y constituye la patología más importante de este cultivo por la severidad de los daños y por la creciente difusión del área geográfica afectada. El empleo de la resistencia genética bajo un manejo integrado de la enfermedad, constituye la estrategia más económica y ambientalmente sustentable para lograr el incremento y la estabilidad en la producción de los cultivos de maíz, reduciendo el uso nocivo de agroquímicos. La reacción a la enfermedad MRC se encuentra influida por una fuerte interacción genotipo-ambiente que dificulta el mejoramiento genético. En los ensayos multiambientales la inconsistencia de la respuesta y la inestabilidad de los genotipos ensayados hace dificultoso llevar a cabo una buena selección basada en los síntomas de la enfermedad. Para contribuir a aumentar la eficacia de los métodos de mejoramiento es posible implementar programas en los que se incluyan herramientas biotecnológicas como los marcadores moleculares, que permiten reducir el efecto ambiental y contribuyen a soslayar, al menos en parte, los inconvenientes planteados por los efectos de la interacción genotipo-ambiente facilitando la identificación de los genotipos buscados. La selección fenotípica realizada convencionalmente, puede ser complementada con la selección asistida por marcadores (marker-assisted selection MAS) consistente en utilizar la información genética que brindan marcadores moleculares de ADN, tales como los SSR, asociados a loci o segmentos cromosómicos (quantitative trait loci QTL) que confieran resistencia a MRC. Si bien los resultados preliminares obtenidos por nuestro grupo de trabajo señalan la presencia de posibles QTLs e informan que la reacción frente a la enfermedad tiene una moderada heredabilidad y una substancial variación debida a la interacción genotipo-ambiente, es necesario confirmar los resultados relativos a los parámetros poblacionales y obtener una mejor delimitación de las regiones genómicas identificadas. Con la finalidad de aumentar la eficiencia de los programas de mejoramiento en el desarrollo de genotipos tolerantes mediante la selección asistida por marcadores utilizando una población de mapeo F2 con un fondo genético diferente y líneas endocriadas recombinantes evaluadas en ensayos multiambientales, se proponen los siguientes objetivos (i) estudiar la forma de herencia de la reacción frente a MRC, (ii) identificar nuevos QTLs asociados a este carácter y (iii) verificar la consistencia de los QTLs identificados previamente.

Estudos experimentais sôbre a origem do milho

1) It may seem rather strange that, in spite of the efforts of a considerable number of scientists, the problem of the origin of indian corn or maize still has remained an open question. There are no fossil remains or archaeological relics except those which are quite identical with types still existing. (Fig. 1). The main difficulty in finding the wild ancestor- which may still exist - results from the fact that it has been somewhat difficult to decide what it should be like and also where to look for it. 2) There is no need to discuss the literature since an excellent review has recently been published by MANGELSDORF and REEVES (1939). It may be sufficient to state that there are basically two hypotheses, that of ST. HILAIRE (1829) who considered Brazilian pod corn as the nearest relative of wild corn still existing, and that of ASCHERSON (1875) who considered Euchlaena from Central America as the wild ancestor of corn. Later hypotheses represent or variants of these two hypotheses or of other concepts, howewer generally with neither disproving their predecessors nor showing why the new hypotheses were better than the older ones. Since nearly all possible combinations of ideas have thus been put forward, it har- dly seems possible to find something theoretically new, while it is essential first to produce new facts. 3) The studies about the origin of maize received a new impulse from MANGELSDORF and REEVES'S experimental work on both Zea-Tripsacum and Zea-Euchlaena hybrids. Independently I started experiments in 1937 with the hope that new results might be obtained when using South American material. Having lost priority in some respects I decided to withold publication untill now, when I can put forward more concise ideas about the origin of maize, based on a new experimental reconstruction of the "wild type". 4) The two main aspects of MANGELSDORF and REEVES hypothesis are discussed. We agree with the authors that ST. HILAIRE's theory is probably correct in so far as the tunicata gene is a wild type relic gene, but cannot accept the reconstruction of wild corn as a homozygous pod corn with a hermaphroditic tassel. As shown experimentally (Fig. 2-3) these tassels have their central spike transformed into a terminal, many rowed ear with a flexible rachis, while possessing at the same time the lateral ear. Thus no explanation is given of the origin of the corn ear, which is the main feature of cultivated corn (BRIEGER, 1943). The second part of the hypothesis referring to the origin of Euchlaena from corn, inverting thus ASCHERSON's theory, cannot be accepted for several reasons, stated in some detail. The data at hand justify only the conclusion that both genera, Euchlaena and Zea, are related, and there is as little proof for considering the former as ancestor of the latter as there is for the new inverse theory. 5) The analysis of indigenous corn, which will be published in detail by BRIEGER and CUTLER, showed several very primitive characters, but no type was found which was in all characters sufficiently primitive. A genetical analysis of Paulista Pod Corn showed that it contains the same gene as other tunicates, in the IV chromosome, the segregation being complicated by a new gametophyte factor Ga3. The full results of this analysis shall be published elsewhere. (BRIEGER). Selection experiments with Paulista Pod Corn showed that no approximation to a wild ancestor may be obtained when limiting the studies to pure corn. Thus it seemed necessary to substitute "domesticated" by "wild type" modifiers, and the only means for achieving this substitution are hybridizations with Euchlaena. These hybrids have now been analysed init fourth generation, including backcrosses, and, again, the full data will be published elsewhere, by BRIEGER and ADDISON. In one present publication three forms obtained will be described only, which represent an approximation to wild type corn. 6) Before entering howewer into detail, some arguments against ST. HILAIRE's theory must be mentioned. The premendelian argument, referring to the instability of this character, is explained by the fact that all fertile pod corn plants are heterozygous for the dominant Tu factor. But the sterility of the homozygous TuTu, which phenotypically cannot be identified, is still unexplained. The most important argument against the acceptance of the Tunicata faetor as wild type relic gene was removed recently by CUTLER (not yet published) who showed that this type has been preserved for centuries by the Bolivian indians as a mystical "medicine". 7) The main botanical requirements for transforming the corn ear into a wild type structure are stated, and alternative solutions given. One series of these characters are found in Tripsacum and Euchlaena : 2 rows on opposite sides of the rachis, protection of the grains by scales, fragility of the rachis. There remains the other alternative : 4 rows, possibly forming double rows of female and male spikelets, protection of kernels by their glumes, separation of grains at their base from the cob which is thin and flexible. 8) Three successive stages in the reconstruction of wild corn, obtained experimentally, are discussed and illustrated, all characterized by the presence of the Tu gene. a) The structure of the Fl hybrids has already been described in 1943. The main features of the Tunicata hybrids (Fig. -8), when compared with non-tunicate hybrids (Fig. 5-6), consist in the absence of scaly protections, the fragility of the rachis and finally the differentiation of the double rows into one male and one female spikelet. As has been pointed out, these characters represent new phenotypic effects of the tunicate factor which do not appear in the presence of pure maize modifiers. b) The next step was observed among the first backcross to teosinte (Fig. 9). As shown in the photography, Fig. 9D, the features are essencially those of the Fl plants, except that the rachis is more teosinte like, with longer internodes, irregular four-row-arrangement and a complete fragility on the nodes. c) In the next generation a completely new type appeared (Fig. 10) which resembles neither corn nor teosinte, mainly in consequence of one character: the rachis is thin and flexible and not fragile, while the grains have an abscission layer at the base, The medium sized, pointed, brownish and hard granis are protected by their well developed corneous glumes. This last form may not yet be the nearest approach to a wild grass, and I shall try in further experiments to introduce other changes such as an increase of fertile flowers per spikelet, the reduction of difference between terminal and lateral inflorescences, etc.. But the nature of the atavistic reversion is alveadwy such that it alters considerably our expectation when looking for a still existing wild ancestor of corn. 9) The next step in our deductions must now consist in an reversion of our question. We must now explain how we may obtain domesticated corn, starting from a hypothetical wild plant, similar to type c. Of the several changes which must have been necessary to attract the attention of the Indians, the following two seem to me the most important: the disappearance of all abscission layers and the reduction of the glumes. This may have been brought about by an accumulation of mutations. But it seems much more probable to assume that some crossing with a tripsacoid grass or even with Tripsacum australe may have been responsible. In such a cross, the two types of abscission layer would be counterbalanced as shown by the Flhybrids of corn, Tripsacum and Euchlaena. Furthermore in later generations a.tu-allele of Tripsacum may become homozygous and substitute the wild tunicate factor of corn. The hypothesis of a hybrid origin of cultivated corn is not completely new, but has been discussed already by HARSHBERGER and COLLINS. Our hypothesis differs from that of MANGELSDORF and REEVES who assume that crosses with Tripsacum are responsible only for some features of Central and North American corn. 10) The following arguments give indirects evidence in support of our hypothesis: a) Several characters have been observed in indigenous corn from the central region of South America, which may be interpreted as "tripsacoid". b) Equally "zeoid" characters seem to be present in Tripsacum australe of central South-America. c) A system of unbalanced factors, combined by the in-tergeneric cross, may be responsible for the sterility of the wild type tunicata factor when homozygous, a result of the action of modifiers, brought in from Tripsacum together with the tuallele. d) The hybrid theory may explain satisfactorily the presence of so many lethals and semilethals, responsible for the phenomenon of inbreeding in cultivated corn. It must be emphasized that corn does not possess any efficient mechanism to prevent crossing and which could explain the accumulation of these mutants during the evolutionary process. Teosinte which'has about the same mechanism of sexual reproduction has not accumulated such genes, nor self-sterile plants in spite of their pronounced preference for crossing. 11) The second most important step in domestication must have consisted in transforming a four rowed ear into an ear with many rows. The fusion theory, recently revived byLANGHAM is rejected. What happened evidently, just as in succulent pXants (Cactus) or in cones os Gymnosperms, is that there has been a change in phyllotaxy and a symmetry of longitudinal rows superimposed on the original spiral arrangement. 12) The geographical distribution of indigenous corn in South America has been discussed. So far, we may distinguish three zones. The most primitive corn appears in the central lowlands of what I call the Central Triangle of South America: east of the Andies, south of the Amazone-Basin, Northwest of a line formed by the rivers São Prancisco-Paraná and including the Paraguay-Basin. The uniformity of the types found in this extremely large zone is astonishing (BRIEGER and CUTLER). To the west, there is the well known Andian region, characterized by a large number of extremely diverse types from small pop corn to large Cuszco, from soft starch to modified sweet corn, from large cylindrical ears to small round ears, etc.. The third region extends along the atlantic coast in the east, from the Caribean Sea to the Argentine, and is characterized by Cateto, an orange hard flint corn. The Andean types must have been obtained very early, and undoubtedly are the result of the intense Inca agriculture. The Cateto type may be obtained easily by crosses, for instance, of "São Paulo Pointed Pop" to some orange soft corn of the central region. The relation of these three South American zones to Central and North America are not discussed, and it seems essential first to study the intermediate region of Ecuador, Colombia and Venezuela. The geograprical distribution of chromosome knobs is rapidly discussed; but it seems that no conclusions can be drawn before a large number of Tripsacum species has been analysed.

Estudo comparativo entre silagem de milho e cana taquara na alimentação suplementar de vacas leiteiras

The A. A. compare corn silage (Zea mays L.) with sugar cane (Saccharum sinensis Roxb.) in the supplementary feeding of dairy cow. Both the feeds were studied in relation to the following points: composition and nutritive value; influence of milk production, milk fat, milk acidity and body weight; cost of production. Both corn silage and sugar cane were analysed by ordinary methods, and their digestibility was determined by means of digestibility coefficients; their composition and nutritive value are, practically, equivalent, but silage showed slight superiority. The feeding experiment was carried out with two groups of six Holstein Friesian cows each, of the Escola Superior de Agricultura "Luiz de Queiroz" herd. Both groups were fed with the same basic concentrates mixture, calculeted according to MORRISON. During the various periods of the experiment, only the roughage supplement varied. The supplementary feeding consisted of 15 kg of chopped sugar cane or corn, silage, per day and per cow, given in two daily meals in the barn. At 4,30 p.m., the cows are set free in the field, where they pass the over night. The experiment was divided into six periods, in which there was a gradative change of the supplementary feeding between the two groups. The milk was weighed every day; the analysis of milk fat and acidity and the weighing of the animals, were made only on the first three days of every week. The analysis of data showed that: a) Milk production was increased significantly by silage feeding; b) The ri was not any influence on milk fat; c) The silage caused higher milk acidity; d) The sugar cane gave a greater increase of body weight. The cost of production of corn silage was 2,12 time higher, than sugar cane, hence, although the silage gave a higher milk production, its use is not economical, compared with sugar cane, in our conditions.

Contribuição para o estudo da composição e valor nutritivo de plantas forrageiras

The authors repport in this paper, the results of trials by means of which were determined the composition and nutritive value of three grasses, that are cultivated in Piracicaba, in State of São Paulo, Brazil: Capim jaragaá, Hyparrhenia rufa, (Nees.) Stapf. (hay); capim de Rhodes, Chloris gayana, Kunth. (hay), and milho Santa Rosa, Zea mays, L. (silage). The digestion trials were carried out with sheep, following the technique of MEAD and GUILBERT. The chemical analysis of the forages were made by methods usually employed. The results are in the tables I and II, of this paper.

Adubação de milho no município de Oliveira, estado de Minas Gerais

Um experimento sobre adubação de milho (Zea maysL.), variedades "Agroceres - 13", foi conduzido no Município de Oliveira, Estado de Minas Gerais, com a finalidade de se estudar o efeito de adubos nitrogenado, fosfatado e potássico sobre a produção desse importante cereal. Os resultados obtidos a partir de um ensaio fatorial 2³ mostraram que a adubação nitrogenada foi a que proporcionou maior produção e renda líquida. Para a adubação fosfatada, houve um pequeno aumento de produção. Não houve resposta ao potássio.

Eficiência de vários adubos fosfatados II. Efeito residual

Estudou-se efeito residual de oito fontes de fósforo em dois solos de baixo teor em fósforo, na cultura de milho (Zea mays L.). Os experimentos foram realizados na casa de vegetação. Pela avaliação do peso de matéria seca e quantidade de nitrogênio, fósforo e potássio absorvidos, constatou-se que os fertilizantes com melhor efeito residual foram metafosfato de cálcio e metafosfato de potássio, seguidos por termofosfato e fosforita de Olinda, no regosol. No latosol não se observou diferenças estatisticamente significativas para peso de materia seca, mas de uma maneira geral o metafosfato de cálcio, fosforita de Olinda e metafosfato de potássio foram os melhores. Os fertilizantes com menor efeito residual foram os que contem alto teor em fósforo solúvel em água: superfosfato triplo, superfosfato simples, fosfato diamonico e polifosfato de amonio.

Ação de reguladores e estimulantes vegetais na germinação de milho e tomateiro

Reguladores e estimulantes vegetais tem sido utilizados nas condições dos trópicos sem a verificação de seus efeitos sob condições controladas, levando frequentemente ao mascaramento de seus efeitos pelas condições do agroecossistema. Com a finalidade de observar os efeitos dessas substâncias sob condições de laboratório, sementes de Zea mays cv. C-525 e Lycopersicon esculentum cv. Kada foram submetidas aos efeitos de giberelina 100 ppm, chlormequat 2000 ppm, hidrazida maleica 1000 ppm, Agrostemin 1,25 g. l-1 e Atonik 0,5 ml.1-1, além do controle, em caixas de Petri. Observações realizadas 7 e 15 dias após a instalação do ensaio mostraram que giberelina e Agrostemin promoveram aumento no crescimento da radícula e do hipocótilo de milho e tomateiro. Hidrazida maleica inibiu o desenvolvimento da radícula das plântulas de milho. Hidrazida maleica e Atonik tenderam a reduzir o crescimento da radícula e do hipocótilo de tomateiro.

Efeitos da aplicação de zinco em solos sob vegetação de cerrado

O presente trabalho teve como objetivo estudar os efeitos da aplicação de zinco em duas unidades de solos originalmente cobertas com vegetação de cerrado, classificadas como Latossolo Vermelho Amarelo-fase arenosa e Areias Quartzosas, que ocorrem no Município de São Simão no Estado de São Paulo Foram feitas determinações químicas do zinco solúvel em HCl 0,1 N, em amostras de solos antes e depois de terem recebido a calagem. As determinações de zinco foram complementadas com o teste microbiológico do Aspergilus niger Simultaneamente, foram conduzidos ensaios de campo com milho (Zea mays L.) e feijão Phaseolus vulgaris L.) para estudar os efeitos da aplicação de zinco nas duas unidades de solo. Outros experimentos com milho foram desenvolvidos sob condições controladas, em casa de vegetação, para estudos de aplicação de zinco. As conclusões do trabalho foram as seguintes: 1. Os resultados dos ensaios microbiológicos mostraram boa concordância com os testes químicos e com a resposta do milho a aplicação de zinco em condições de campo. 2. A cultura do milho deu boas respostas a aplicação de zinco quando cultido nos solos Areias Quartzosas, correspondendo a uma dose econômica de 2,4 kg/ha de zinco. 3. Não houve efeito do zinco para o milho quando este foi cultivado no Latossolo Vermelho Amarelo-fase arenosa. 4. Não houve efeitos do zinco na produção do feijoeiro nas duas unidades de solos. 5. O ensaio de vasos usando o milho como planta teste não foi eficiente na resposta dos solos as aplicações de zinco.

Efeitos do triacontanol na germinação de milho e tomateiro

Novos estimulantes vegetais têm merecido estudos por possuírem alto potencial para melhorarem a produtividade de culturas. Triacontanol, um álcool alifático primário de cadeia longa isolado de vegetais encontra-se dentre essas substâncias. Com o objetivo de verificar o efeito deste estimulante vegetal sob condições de laboratório, sementes de Zea mays cv. C-525 e Lycopersicon esculentum cv. Kada foram submetidas aos efeitos de 10 ml de triacontanol 0,01, 0,1 e 1 mg. 1-1, além do controle, em caixas de Petri. Observações realizadas 7 e 14 dias após a instalação do ensaio mostraram que concentrações crescentes de triacontanol tendem a aumentar o crescimento da radícula e do hipocótilo do tomateiro, 14 dias após a aplicação. Nesta data notou-se também que concentrações crescentes de triacontanol tendem a incrementar o desenvolvimento do hipocótilo do milho.

Nutrição mineral de hortaliças: LXXXIX. Crescimento e acúmulo de macro e micronutrientes por uma cultura de milho doce

De uma plantação bem conduzida de milho doce (Zea mays var. saahirata) cv. Contibrasil situada sobre um Latossolo Vermelho Escuro Orto, série "Luiz de Queiroz" de alta fertilidade natural no município de Piracicaba, SP, foram coletadas plantas aos 45, 60, 75, 90 e 105 dias após a germinação. As plantas foram divididas em folhas novas, velhas, colmo, pendão e espigas. O material após sofrer os processos necessários foi seco, pesado e analisado para N, P, K, Ca, Mg, S, B, Cu, Fe, Mn e Zn por métodos convencionais de laboratório. Os autores concluíram que: aumento de peso de matéria seca total é contínua até o final do ciclo; a acumulação de macronutrientes pela planta inteira obedece a seguinte ordem decrescente: N, K, P,Ca=S e Mg; o maior acúmulo de macronutrientes no final do ciclo ocorre no colmo, com exceção do N que é acumulado em maior quantidade nas espigas; o acúmulo de micronutrientes pela planta inteira obedece a seguinte ordem decrescente: Fe, Mn, Cu, Zn e B; o maior acúmulo de micronutrientes no final do ciclo ocorre no colmo; a exportação de nutrientes pela espiga representa 26,15% do total de nutrientes contidos na plantação.

Ablation of caterpillar labial salivary glands: technique for determining the role of saliva in insect-plant interactions.

There has been an ardent interest in herbivore saliva due to its roles in inducing plant defenses and its impact on herbivore fitness. Two techniques are described that inhibit the secretion of labial saliva from the caterpillar, Helicoverpa zea, during feeding. The methods rely on cauterizing the caterpillar's spinneret, the principal secretory structure of the labial glands, or surgically removing the labial salivary gland. Both methods successfully inhibit secretion of saliva and the principal salivary enzyme glucose oxidase. Caterpillars with inhibited saliva production feed at similar rates as the untreated caterpillars, pupate, and emerge as adults. Glucose oxidase has been suggested to increase the caterpillar's survival through the suppression of inducible anti-herbivore defenses in plants. Tobacco (Nicotiana tabacum) leaves fed on by caterpillars with ablated salivary glands had significantly higher levels of nicotine, an inducible anti-herbivore defense compound of tobacco, than leaves fed upon by caterpillars with intact labial salivary glands. Tomato (Lycopersicon esculentum) leaves fed upon by caterpillars with suppressed salivary secretions showed greatly reduced evidence of hydrogen peroxide formation compared to leaves fed upon by intact caterpillars. These two methods are useful techniques for determining the role that saliva plays in manipulating plant anti-herbivore defenses.

Use of the pyrophosphatase activity as a reliable tonoplast marker in maize roots.

Membranes of maize (Zea mays L., cv LG 11) roots were fractionated by sucrose (in presence or absence of Mg2+) or dextran density gradient centrifugations and the locations of organelles were determined using marker enzymes. Latent UDPase was used as a Golgi marker, catalase for the peroxysomes, cytochrome c oxidase for the mitochondria, UDP-Gal-galactosyltransferase for the amyloplast membranes and NADH-cytochrome c reductase for the ER. Two markers were selected for the plasmalemma, the vanadate-sensitive ATPase and UDP-Glc-sterolglucosyltransferase. The distributions of the PPase and vacuolar ATPase were found to be similar after density gradient centrifugation. The PPase and vacuolar ATPase activities were clearly separated from almost all the other markers tested, however, a partial association of both activities with the ER cannot be completely ruled out. The PPase of maize roots is more active and easier to measure than the vacuolar ATPase and is therefore an excellent candidate for use as a tonoplast marker.

A Ca2+/H+ antiport system driven by the tonoplast pyrophosphate-dependent proton pump from maize roots.

Calcium uptake by tonoplast enriched membrane vesicles from maize (Zea mays L. cv. LG 11) primary roots was studied. A pH gradient, measured by the fluorescence quenching of quinacrine, was generated across sealed vesicles driven by the pyrophosphate-dependent proton pump. The fluorescence quenching was strongly inhibited by Ca2+; moreover, when increasing Ca2+ concentrations were added to vesicles at steady-state, a concomitant decrease in the proton gradient was observed. Ca2+ uptake using Ca-45(2+) was linear from 10 min when oxalate (10 mM) was present, while Ca2+ uptake was completely inhibited with proton ionophores (FCCP and monensin), indicating a Ca2+/H+ antiport. Membranes were further fractionated using a linear sucrose density gradient (10-45%) and were identified with marker enzymes. Ca2+ uptake co-migrated with the tonoplast pyrophosphate-dependent proton pumping, pyrophosphatase and ATPase activities: the Ca2+/H+ antiport is consequently located at the tonoplast.