Thylakoid membrane architecture

Confocal scanning laser microscopic observations were made on live chloroplasts in intact cells and on mechanically isolated, intact chloroplasts. Chlorophyll fluorescence was imaged to observe thylakoid membrane architecture. C-3 plant species studied included Spinacia oleracea L., Spathiphyllum sp. Schott, cv. 'Mauna Loa', and Pisum sativum L. C-4 plants were also investigated: Saccharum officinarum L., Sorghum bicolor L. Moench, Zea mays L. and Panicum miliaceum L. Some Spinacia chloroplasts were treated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) to enhance or sodium dithionite (SD) to reduce the photosystem II fluorescence signal. Confocal microscopy images of C-3 chloroplasts differed from electron microscopy pictures because they showed discrete spots of bright fluorescence with black regions between them. There was no evidence of fluorescence from stroma thylakoids. The thylakoid membrane system at times appeared to be string-like, with brightly fluorescing grana lined up like beads. C-4 bundle sheath chloroplasts were imaged from three different types of C-4 plants. Saccharum and Sorghum bundle sheath chloroplasts showed homogeneous fluorescence and were much dimmer than mesophyll chloroplasts. Zea had rudimentary grana, and dim, homogeneous intergrana fluorescence was visualised. Panicum contained thylakoids similar in appearance and string-like arrangement to mesophyll chloroplasts. Isolated Pisum chloroplasts, treated with a drop of 5 mM MgCl2 showed a thylakoid membrane system which appeared to be unravelling. Spongy mesophyll chloroplasts of Spinacia treated with 5 mM sodium dithionite showed a granal thylakoid system with distinct regions of no fluorescence. A time-series experiment provided evidence of dynamic membrane rearrangements over a period of half an hour.

Long-distance signalling and a mutational analysis of branching in pea

Four ramosus mutants with increased branching at basal and aerial nodes have been used to investigate the genetic regulation of bud outgrowth in Pisum sativum L. (garden pea). Studies of long-distance signalling, xylem sap cytokinin concentrations, shoot auxin level, auxin transport and auxin response are discussed. A model of branching control is presented that encompasses two graft-transmissible signals in addition to auxin and cytokinin. Mutants rms1 through rms4 are not deficient in indole-3-acetic acid (IAA) or in the basipetal transport of this hormone. Three of the four mutants, rms1, rms3 and rms4, have very reduced cytokinin concentrations in xylem sap from roots. This reduction in xylem sap cytokinin concentration appears to be caused by a property of the shoot and may be part of a feedback mechanism induced by an aspect of bud outgrowth. The shoot-to-root feedback signal is unlikely to be auxin itself, as auxin levels and transport are not correlated with xylem sap cytokinin concentrations in various intact and grafted mutant and wild-type plants. Rms1 and Rms2 act in shoot and rootstock to regulate the level or transport of graft-transmissible signals. Various grafting studies and double mutant analyses have associated Rms2 with the regulation of the shoot-to-root feedback signal. Rms1 is associated with a second unknown graft-transmissible signal that is postulated to move in the direction of root-to-shoot. Exogenous auxin appears to interact with both of the signals regulated by Rms1 and Rms2 in the inhibition of branching after decapitation. The action of Rms3 and Rms4 is less apparent at this stage, although both appear to act largely in the shoot.

The shoot controls zeatin riboside export from pea roots. Evidence from the branching mutant rms4

The rms4 mutant of pea (Pisum sativum L.) was used in grafting studies and cytokinin analyses of the root xylem sap to provide evidence that, at least for pea, the shoot can modify the import of cytokinins from the root. The rms4 mutation, which confers a phenotype with increased branching in the shoot, causes a very substantial decrease (down to 40-fold less) in the concentration of zeatin riboside (ZR) in the xylem sap of the roots. Results from grafts between wild-type (WT) and rms4 plants indicate that the concentration of cytokinins in the xylem sap of the roots is determined almost entirely by the genotype of the shoot. WT scions normalize the cytokinin concentration in the sap of rms4 mutant roots, whereas mutant scions cause WT roots to behave like those of self-grafted mutant plants. The mechanism whereby rms4 shoots of pea cause a down-regulation in the export of cytokinins from the roots is unknown at this time. However, our data provide evidence that the shoot transmits a signal to the roots and thereby controls processes involved in the regulation of cytokinin biosynthesis in the root.

The rms1 mutant of pea has elevated indole-3-acetic acid levels and reduced root-sap zeatin riboside content but increased branching controlled by graft-transmissible signal(s)

Rms1 is one of the series of five ramosus loci in pea (Pisum sativum L.) in which recessive mutant alleles confer increased branching at basal and aerial vegetative nodes. Shoots of the nonallelic rms1 and rms2 mutants are phenotypically similar in most respects. However, we found an up to 40-fold difference in root-sap zeatin riboside ([9R]Z) concentration between rms1 and rms2 plants. Compared with wild-type (WT) plants, the concentration of [9R]Z in rms1 root sap was very low and the concentration in rms2 root sap was slightly elevated. To our knowledge, the rms1 mutant is therefore the second ramosus mutant (rms4 being the first) to be characterized with low root-sap [9R]Z content. Like rms2, the apical bud and upper nodes of rms1 plants contain elevated indole-3-acetic acid levels compared with WT shoots. Therefore, the rms1 mutant demonstrates that high shoot auxin levels and low root-sap cytokinin levels are not necessarily correlated with increased apical dominance in pea. A graft-transmissible basis of action has been demonstrated for both mutants from reciprocal grafts between mutant and WT plants. Branching was also largely inhibited in rms1 shoots when grafted to rms2 rootstocks, but was not inhibited in rms2 shoots grafted to rms1 rootstocks. These grafting results are discussed, along with the conclusion that hormone-like signals other than auxin and cytokinin are also involved.

Hormone physiology of pea mutants prevented from flowering by mutations gi or veg1

The veg1 (vegetative) mutant in pea (Pisum sativum L.) does not flower under any circumstances and gi (gigas) mutants remain vegetative under certain conditions. gi plants are deficient in production of floral stimulus, whereas veg1 plants lack a response to floral stimulus. During long days in particular, these non-flowering mutant plants eventually enter a stable compact phase characterised by a large reduction in internode length, small leaves and growth of lateral shoots from the upper-stem (aerial) nodes. The first-order laterals in turn produce second-order laterals and so on in a reiterative pattern. The apical bud is reduced in size but continues active growth. Endogenous hormone measurements and gibberellin application studies with gi-1, gi-2 and veg1 plants indicate that a reduction in gibberellin and perhaps indole-3-acetic acid level may account, at least partially, for the compact aerial shoot phenotype. In the gi-1 mutant, the compact phenotype is rescued by transfer from a 24- to an 8-h photoperiod. We propose that in plants where flowering is prevented by a lack of floral stimulus or an inability to respond, the large reduction in photoperiod gene activity during long days may lead to a reduction in apical sink strength that is manifest in an altered hormone profile and weak apical dominance.

Pea rms6 mutants exhibit increased basal branching

Our studies on two branching mutants of pea (Pisum sativum L.) have identified a further Ramosus locus, Rms6, with two recessive or partially recessive mutant alleles: rms6-1 (type line S2-271) and rms6-2 (type line K586). Mutants rms6-1 and rms6-2 were derived from dwarf and tall cultivars, Solara and Torsdag, respectively. The rms6 mutants are characterized by increased branching from basal nodes. In contrast, mutants rms1 through rms5 have increased branching from both basal and aerial (upper stem) nodes. Buds at the cotyledonary node of wild-type (WT) plants remain dormant but in rms6 plants these buds were usually released from dormancy. Their growth was either subsequently inhibited, sometimes even prior to emergence above ground, or they grew into secondary stems. The mutant phenotype was strongest for rms6-1 on the dwarf background. Although rms6-2 had a weak single-mutant phenotype, the rms3-1 rms6-2 double mutant showed clear transgression and an additive branching phenotype, with a total lateral length almost 2-fold greater than rms3-1 and nearly 5-fold greater than rms6-2 . Grafting studies between WT and rms6-1 plants demonstrated the primary action of Rms6 may be confined to the shoot. Young WT and rms6-1 shoots had similar auxin levels, and decapitated plants had a similar magnitude of response to applied auxin. Abscisic acid levels were elevated 2-fold at node 2 of young rms6-1 plants. The Rms6 locus mapped to the R to Gp segment of linkage group V (chromosome 3). The rms6 mutants will be useful for basic research and also have possible agronomical value.

Leaf area development of ABA-deficient and wild-type peas at two levels of nitrogen supply

The ABA-deficient wilty pea (Pisum sativum L.) and its wild-type (WT) were grown at two levels of nitrogen supply (0.5 and 5.0 mM) for 5-6 weeks from sowing, to determine whether leaf ABA status altered the leaf growth response to N deprivation. Plants were grown at high relative humidity to prevent wilting of the wilty peas. Irrespective of N supply, expanding wilty leaflets had ca 50% less ABA than WT leaflets but similar ethylene evolution rates. Fully expanded wilty leaflets had lower relative water contents (RWC) and were 10-60% smaller in area (according to the node of measurement) than WT leaflets. However, there were no genotypic differences in plant relative leaf expansion rate (RLER). Growth of both genotypes at 0.5 mM N increased the RWC of fully expanded leaflets, but did not alter ethylene evolution or ABA concentration of expanding leaflets. Plants grown at 0.5 mM N showed a 20-30% reduction in RLER, which was similar in magnitude in both wilty and WT peas. Thus, leaf ABA status did not alter the leaf growth response to N deprivation.

Mineralização do nitrogênio de plantas de cobertura, solteiras e consorciadas, depositadas sobre um solo com histórico de cultivo de cebola

A decomposição das plantas de cobertura de inverno, depositadas sobre o solo, podem aumentar a disponibilidade de formas de nitrogênio (N), durante o ciclo da cebola. O trabalho objetivou avaliar a mineralização de N da massa de plantas de cobertura, solteiras e consorciadas, em um solo com histórico de cultivo de cebola. Porções de solo foram coletadas, preparadas, acondicionadas em recipientes de acrílico. Matéria seca de aveia preta, centeio, nabo-forrageiro, aveia preta+nabo-forrageiro e centeio+nabo-forrageiro, foram adicionadas sobre a superfície do solo e incubadas por 90 dias. No tempo zero e aos 18, 36, 54, 72 e 90 dias após a incubação (DAI), as porções de solo foram amostradas e determinados os teores de N total, N-NO3- e N-NH4+ de cada uma. Calcularam-se os valores de N mineral, N mineral líquido, N mineralizado e N total-N mineralizado. Os maiores teores de N-NH4+ foram observados nas porções de solo com a deposição de massa de nabo-forrageiro e do consórcio centeio+nabo-forrageiro. Os maiores teores de N-NO3- e N-mineral dos 36 até os 90 DAI e de N-mineralizado dos 18 até os 92 DAI foram observados nas porções de solo com a deposição de massa de centeio + nabo-forrageiro. A taxa de mineralização foi positiva em todas as amostras do solo com deposição de massa de centeio e nabo-forrageiro, e dos consórcios aveia preta+nabo-forrageiro e centeio+nabo-forrageiro e negativa aos 18 e 72 DAI, nas porções de solo com deposição de massa de aveia. Os resíduos de nabo-forrageiro e do consórcio centeio+nabo-forrageiro apresentaram o maior potencial de mineralização.

Atratividade de adultos de Chrysoperla externa (Hagen, 1861) aos compostos voláteis de coentro, endro e erva-doce (Apiaceae) em condições de laboratório

Espécies de Apiaceae dispõem de óleos essenciais, nos quais podem ocorrer compostos voláteis, que funcionam como sinais para atração e manutenção de inimigos naturais nas áreas cultivadas. Com base nestas características, este trabalho objetivou avaliar a atratividade aos adultos do predador Chrysoperla externa. Foram utilizados folhas e caules de coentro, endro e erva-doce, coletados aos 30 e 60 dias após a semeadura. As plantas foram dispostas em olfatômetro de quatro vias (formato de "X") disponibilizando-se os odores para machos e fêmeas, virgens e acasalados, em testes de livre escolha. Ao serem liberados individualmente no interior do olfatômetro, foram cronometrados cinco minutos e contabilizado o tempo total de permanência do inseto em cada braço do aparelho. Os dados foram analisados pelo teste c², com frequência esperada de 25%. Estudou-se o rendimento de óleo essencial das três espécies de plantas, 30 e 60 dias após a semeadura, utilizando-se do método de hidrodestilação. A composição química dos óleos foi determinada por cromatografia gasosa acoplada a espectrômetro de massas. Verificou-se que adultos virgens têm preferência por plantas de coentro, enquanto os acasalados preferem plantas de erva-doce, ambas coletadas aos 30 dias. Plantas com 60 dias não proporcionaram resposta atrativa aos adultos de C. externa. O rendimento de óleo tendeu a aumentar com o desenvolvimento fenológico da planta. A composição química do óleo de coentro revelou, como componentes majoritários, o (2E)-decenal e decanal e, para erva-doce, a maior concentração foi de (E)-anetol.

Crescimento e nutrição mineral de cebolinha verde cultivada hidroponicamente sob diferentes concentrações de N, P e K

RESUMO Em cultivos hidropônicos de hortaliças folhosas destacam-se, dentre os fatores que influenciam a produção e qualidade do produto, as exigências nutricionais da cultura. Objetivou-se, com este trabalho, avaliar o efeito de concentrações de N, P e K sobre a produção de matéria seca e a nutrição mineral em cebolinha 'Todo ano' (Allium fistulosum L.), cultivada em condições hidropônicas. Foram conduzidos três experimentos, um com cada nutriente em três níveis: deficiente, adequado e excessivo. Empregou-se o delineamento inteiramente casualizado, em esquema de parcelas subdivididas no tempo, com quatro repetições e dez plantas por parcela. Aos 15, 30, 45 e 60 dias após o transplante (DAT), foi avaliada a produção de matéria seca da parte aérea e de raízes. Aos 45 e 60 DAT, avaliaram-se os teores e os acúmulos totais de N, P e K e os teores de Ca e Mg da parte aérea da planta. Em cultivos hidropônicos, as deficiências de N e P são mais limitantes que a de K ao crescimento da cebolinha cultivar 'Todo Ano', sendo que níveis baixos de P afetam principalmente o crescimento do sistema radicular da planta. A deficiência de N afeta a absorção de P, K, Ca e Mg. O excesso de N em cebolinha manifesta-se pelo crescimento excessivo da parte aérea e pelo aumento da flacidez das folhas. Os excessos de P e de K na solução nutritiva não provocam sintomas visuais de toxicidade de P ou de K, mas níveis excessivos de K diminuem os teores de Ca e de Mg da parte aérea da planta.

Efeito de doses de fósforo na produtividade e armazenamento pós-colheita de dois cultivares de cebola

RESUMO O fósforo é, reconhecidamente, o nutriente-chave para a obtenção de produtividades elevadas e tem sido o macronutriente que mais frequentemente limita a produção, em solos pobres nesse nutriente. Com o objetivo de avaliar a produtividade e armazenamento pós-colheita de bulbos de cultivares de cebola, em função de doses de fósforo, conduziu-se um experimento, de maio a dezembro de 2011, em Petrolina-PE. O delineamento experimental utilizado foi de blocos ao acaso, no esquema fatorial 5x2, compreendendo cinco doses de fósforo (0; 60; 120, 180 e 240 kg ha-1) e dois cultivares (Franciscana IPA-10 e Vale Ouro IPA-11), com quatro repetições. Os cultivares Franciscana IPA-10 (74,6 t ha-1) e Vale Ouro IPA-11 (76,1 t ha-1) não apresentaram diferenças significativas para produtividade comercial de bulbos. Maior produtividade foi obtida na dose de 132 kg ha-1 de P2O5, associada à dose mais econômica de 130 kg ha-1 de P2O5. Verificou-se com o aumento das doses uma redução gradativa da produção de bulbos considerados não comerciais (refugos), sendo a menor produção estimada de refugos a obtida com a dose de 124 kg ha-1 de P2O5. Bulbos comerciais com maiores massa fresca e diâmetro foram obtidos com o incremento das doses. Não se verificou perda de massa significativa aos 30 dias após cura, para doses ou cultivares. Aos 60 dias após cura, detectou-se efeito significativo com menor perda para o cultivar Franciscana IPA-10 (27,2%) em comparação com o Vale Ouro IPA-11 (31,9%).

Herbicidal activity of volatiles from coriander, winter savory, cotton lavender, and thyme isolated by hydrodistillation and supercritical fluid extraction

The volatiles from Coriandrum sativum L., Satureja montana L., Santolina chamaecyparissus L., and Thymus vulgaris L. were isolated by hydrodistillation (essential oil) and supercritical fluid extraction (volatile oil). Their effect on seed germination and root and shoot growth of the surviving seedlings of four crops (Zea mays L., Triticum durum L., Pisum sativum L., and Lactuca sativa L.) and two weeds (Portulaca oleracea L. and Vicia sativa L.) was investigated and compared with those of two synthetic herbicides, Agrocide and Prowl. The volatile oils of thyme and cotton lavender seemed to be promising alternatives to the synthetic herbicides because they were the least injurious to the crop species. The essential oil of winter savory, on the other hand, affected both crop and weeds and can be appropriate for uncultivated fields.

Extraction of volatile oil from aromatic plants with supercritical carbon dioxide: experiments and modeling

An overview of the studies carried out in our laboratories on supercritical fluid extraction (SFE) of volatile oils from seven aromatic plants: pennyroyal (Mentha pulegium L.), fennel seeds (Foeniculum vulgare Mill.), coriander (Coriandrum sativum L.), savory (Satureja fruticosa Beguinot), winter savory (Satureja montana L.), cotton lavender (Santolina chamaecyparisus) and thyme (Thymus vulgaris), is presented. A flow apparatus with a 1 L extractor and two 0.27 L separators was built to perform studies at temperatures ranging from 298 to 353 K and pressures up to 30.0 MPa. The best compromise between yield and composition compared with hydrodistillation (HD) was achieved selecting the optimum experimental conditions of extraction and fractionation. The major differences between HD and SFE oils is the presence of a small percentage of cuticular waxes and the relative amount of thymoquinone, an oxygenated monoterpene with important biological properties, which is present in the oils from thyme and winter savory. On the other hand, the modeling of our data on supercritical extraction of volatile oil from pennyroyal is discussed using Sovova's models. These models have been applied successfully to the other volatile oil extractions. Furthermore, other experimental studies involving supercritical CO2 carried out in our laboratories are also mentioned.

Contribuição ao estudo das plantas medicinais no Estado de Alagoas V

O nosso trabalho é mais uma etapa, a quinta, do estudo sobre as plantas medicinais, ocorrentes no Estado de Alagoas. Várias excursões foram feitas, a fim de coletar espécies, utilizadas na medicina caseira. Os espécimens coletados foram preparados, identificados e incorporados ao Herbario "Professor Honorio Monteiro" (MUFAL) da Universidade Federal de Alagoas. O estudo das plantas medicinais, abrangendo famílias, nomes científicos e vulgares e a descrição sucinta para cada espécie, bem como o uso medicinal e a posologia, foi feito com auxílio de bibliografias especializadas no assunto. Foram estudadas as seguintes espécies, consideradas medicinais: Allium ascalonicum L. (cebola-branca); Astronium urundeuva (Fr. All.) Engl. (aroeira-do-sertão); Cecropia sp. (imbáuba); Coix lacryma-jobi L. (capim-de-contas); Daucus carota L. (cenoura); Eucalyptus citriodora Hook. (eucalípto); Eugenia jambosa L. (jambo-rosa); Eugenia malaccensis L. (jambo-roxo); Genipa americana L. (genipapo); Guazuma ulmifolia Lam. (mutamba); Helianthus annuusL. (girassol); Hedychium coronarium Koening (lírio-do-brejo); Imperata brasiliensis Trin. (capim-sapê); Jatropha curcas L. (pinhão-manso) Melocactus bahiensis (Brit. et Rose) Luetzelb. (coroa-de-frade), Monniera trifolia L. (alfavaca-de-cobra), Pithecollobium avaremotemo Mart. (bordão-de-velho); Polygonum persicaria L. (erva-de-bicho); Solidago microglossa DC. (erva-lanceta) e Syzygium jambolana DC. (azeitona).