The knock-out of ARP3a gene affects F-actin cytoskeleton organization altering cellular tip growth, morphology and development in moss Physcomitrella patens.

The seven subunit Arp2/3 complex is a highly conserved nucleation factor of actin microfilaments. We have isolated the genomic sequence encoding a putative Arp3a protein of the moss Physcomitrella patens. The disruption of this ARP3A gene by allele replacement has generated loss-of-function mutants displaying a complex developmental phenotype. The loss-of function of ARP3A gene results in shortened, almost cubic chloronemal cells displaying affected tip growth and lacking differentiation to caulonemal cells. In moss arp3a mutants, buds differentiate directly from chloronemata to form stunted leafy shoots having differentiated leaves similar to wild type. Yet, rhizoids never differentiate from stem epidermal cells. To characterize the F-actin organization in the arp3a-mutated cells, we disrupted ARP3A gene in the previously described HGT1 strain expressing conditionally the GFP-talin marker. In vivo observation of the F-actin cytoskeleton during P. patens development demonstrated that loss-of-function of Arp3a is associated with the disappearance of specific F-actin cortical structures associated with the establishment of localized cellular growth domains. Finally, we show that constitutive expression of the P. patens Arp3a and its Arabidopsis thaliana orthologs efficiently complement the mutated phenotype indicating a high degree of evolutionary conservation of the Arp3 function in land plants.

HydroxyprolineO-arabinosyltransferase mutants oppositely alter tip growth inArabidopsis thalianaandPhyscomitrella patens

Hydroxyproline O-arabinosyltransferases (HPATs) are members of a small, deeply conserved family of plant-specific glycosyltransferases that add arabinose sugars to diverse proteins including cell wall-associated extensins and small signaling peptides. Recent genetic studies in flowering plants suggest that different HPAT homologs have been co-opted to function in diverse species-specific developmental contexts. However, nothing is known about the roles of HPATs in basal plants. We show that complete loss of HPAT function in Arabidopsis thaliana and the moss Physcomitrella patens results in a shared defect in gametophytic tip cell growth. Arabidopsis hpat1/2/3 triple knockout mutants suffer from a strong male sterility defect as a consequence of pollen tubes that fail to fully elongate following pollination. Knocking out the two HPAT genes of Physcomitrella results in larger multicellular filamentous networks due to increased elongation of protonemal tip cells. Physcomitrella hpat mutants lack cell-wall associated hydroxyproline arabinosides and can be rescued with exogenous cellulose, while global expression profiling shows that cell wall-associated genes are severely misexpressed, implicating a defect in cell wall formation during tip growth. Our findings point to a major role for HPATs in influencing cell elongation during tip growth in plants.

Polarized cell growth in Arabidopsis requires endosomal recycling mediated by GBF1-related ARF exchange factors.

Polarized tip growth is a fundamental cellular process in many eukaryotic organisms, mediating growth of neuronal axons and dendrites or fungal hyphae. In plants, pollen and root hairs are cellular model systems for analysing tip growth. Cell growth depends on membrane traffic. The regulation of this membrane traffic is largely unknown for tip-growing cells, in contrast to cells exhibiting intercalary growth. Here we show that in Arabidopsis, GBF1-related exchange factors for the ARF GTPases (ARF GEFs) GNOM and GNL2 play essential roles in polar tip growth of root hairs and pollen, respectively. When expressed from the same promoter, GNL2 (in contrast to the early-secretory ARF GEF GNL1) is able to replace GNOM in polar recycling of the auxin efflux regulator PIN1 from endosomes to the basal plasma membrane in non-tip growing cells. Thus, polar recycling facilitates polar tip growth, and GNL2 seems to have evolved to meet the specific requirement of fast-growing pollen in higher plants.

Influência da convecção termossolutal nos espaçamentos dendríticos primários durante a solidificação direcional horizontal de ligas Sn-Pb

Este trabalho apresenta um estudo teórico-experimental sobre os efeitos da convecção termossolutal sobre os espaçamentos dendríticos primários de ligas hipoeutéticas Sn-Pb. Assim, um dispositivo experimental de configuração horizontal refrigerado a água foi construído e amostras do referido sistema de ligas foram solidificadas direcionalmente sob condições transientes de extração de calor. Um método teórico-experimental é aplicado para determinar as velocidades de deslocamento da isoterma liquidus, os gradientes térmicos e as taxas de resfriamento. São também apresentados resultados para os respectivos coeficientes de transferência de calor na interface metal/molde os quais foram calculados a partir de uma análise comparativa realizada entre os perfis experimentais de temperatura e valores teóricos fornecidos por um método numérico baseado em volumes finitos. Alguns modelos teóricos para a previsão dos espaçamentos dendríticos primários são comparados com os resultados experimentais obtidos. Finalmente, um estudo comparativo é realizado entre resultados encontrados neste trabalho e aqueles apresentados na literatura para os espaçamentos dendríticos primários das ligas Sn-Pb investigadas quando solidificadas direcionalmente nos sistemas verticais ascendente e descendente, sob as mesmas condições. A análise das microestruturas indica que os espaçamentos dendríticos primários são bastante influenciados pela direção de crescimento do sólido.

Influência da convecção termossolutal na transição colunar/equiaxial em ligas Al-Si sob condições unidirecionais e transitórias de extração de calor

A relação entre macroestrutura e propriedades mecânicas de um material tem sido objeto de intensa investigação pois o tamanho dos grãos, a orientação cristalina e a distribuição dos mesmos exercem influência direta no comportamento mecânico dos produtos acabados. Assim, o entendimento dos fatores que influenciam a formação das zonas estruturais coquilhada, colunar e equiaxial nos materiais fundidos como, por exemplo, o sistema de liga, composição da liga, temperatura de vazamento, temperatura do molde, material do molde, coeficientes de transferência de calor na interface metal/molde, taxa de resfriamento, gradientes térmicos, dimensão da peça, presença de convecção no líquido, transporte de soluto, etc é de fundamental importância para a melhoria da eficiência do processo de fundição envolvido. Com base no conhecimento dos princípios termofísicos em que essas zonas são formadas, é possível manipular de forma bastante razoável a estrutura dos produtos fundidos e, conseqüentemente, as propriedades mecânicas dos mesmos. Tendo como principal foco a análise da mudança da zona colunar para a equiaxial, este trabalho apresenta um estudo teórico-experimental sobre a transição colunar/equiaxial (TCE) das ligas hipoeutéticas Al- 3%Si, Al-7%Si Al-9%Si solidificadas unidirecionalmente em um dispositivo horizontal refrigerado a água sob condições transientes de fluxo de calor. A condição de contato térmico na superfície de extração de calor foi padronizada como sendo polida. Os perfis térmicos foram medidos em diferentes posições do lingote e os dados foram armazenados automaticamente. Um método numérico é utilizado na determinação de variáveis térmicas de solidificação como coeficientes de transferência de calor na interface metal/molde (h_i), velocidades das isotermas liquidus (V_L), gradientes térmicos (G_L) e taxas de resfriamento (T_R) que influenciam diretamente a referida transição estrutural. Os resultados teóricos e experimentais apresentaram boa concordância. Um estudo comparativo entre os resultados obtidos neste trabalho e valores propostos na literatura para analisar a TCE durante a solidificação unidirecional vertical ascendente sob condições transientes de extração de calor das ligas investigadas, também é apresentado.

Influência dos parâmetros térmicos de solidificação na transição colunar/equiaxial em ligas do sistema Sn-Pb sob condições transitórias de extração de calor

O principal objetivo deste trabalho é desenvolver um estudo teórico/experimental sobre a influência dos parâmetros térmicos de solidificação (V _L e T) na transição colunar/equiaxial das ligas Sn 5%Pb, Sn 15%Pb, Sn 20%Pb e Sn 25%Pb sob condições de solidificação unidirecional horizontal refrigerada à água. Inicialmente, são calculadas experimentalmente as velocidades da isoterma líquidus e as taxas de resfriamento das ligas em questão cujos resultados são comparados com as previsões teóricas de um modelo numérico. Em seguida, a posição da transição colunar/equiaxial é determinada por meio da análise macroestrutural assim como são avaliados os efeitos impostos por correntes convectivas devido ao efeito do soluto no comportamento da transição colunar/equiaxial das referidas ligas. Finalmente, é realizado um estudo experimental comparativo para as ligas estudadas quando solidificadas unidirecionalmente em diferentes sistemas refrigerados à água.

Influência das variáveis térmicas sobre os espaçamentos dendríticos terciários durante a solidificação direcional horizontal da liga Al-6%Cu

A liga Al-6%Cu foi solidificada direcionalmente sob condições transitórias de extração de calor e microestruturas dendríticas, variáveis térmicas de solidificação, ou seja, velocidade de deslocamento da isoterma líquidus (V_L), taxa de resfriamento (T_R) e gradiente de temperatura à frente da isoterma liquidus (G_L) foram caracterizadas, determinadas experimentalmente e correlacionadas com os espaçamentos dentríticos terciários (λ₃). Para tanto, foi projetado, construído e aferido um dispositivo de solidificação direcional horizontal. Os resultados encontrados mostram que leis de potência -1,1 e -0,55 caracterizam a variação dos espaçamentos terciários com a velocidade de deslocamento da isoterma liquidus (V_L) e a taxa de resfriamento (T_R), respectivamente. Finalmente, é realizado um estudo comparativo entre os resultados obtidos neste trabalho e aqueles publicados na literatura para ligas Al-Cu solidificadas direcionalmente sob condições transientes de fluxo de calor nos sistemas verticais ascendente e descendente.

Espaçamentos dendríticos primários da liga Sn-5%Pb solidificada direcionalmente em um sistema horizontal

Este trabalho desenvolve um estudo teórico-experimental sobre a correlação entre parâmetros térmicos e os espaçamentos dendríticos primários da liga Sn-5%Pb solidificada direcionalmente em um sistema horizontal sob condições transientes de extração de calor. Os perfis de temperatura foram medidos em diferentes posições do lingote e os dados obtidos foram armazenados automaticamente. São apresentados resultados para o coeficiente de transferência de calor na interface metal/molde o qual foi calculado a partir de uma análise comparativa entre os perfis térmicos experimentais e valores teóricos fornecidos por um modelo numérico. Um método teórico-experimental é aplicado para determinar as velocidades de deslocamento da isoterma liquidus e as taxas de resfriamento. Os resultados teóricos e experimentais levantados apresentaram boa concordância. Um estudo comparativo é realizado entre os dados encontrados neste trabalho e aqueles apresentados na literatura para os espaçamentos dendríticos primários da liga investigada quando solidificada direcionalmente nos sistemas verticais ascendente e descendente, sob as mesmas condições assumidas neste trabalho. A análise das microestruturas indica que os espaçamentos dendríticos primários são bastante influenciados pelos parâmetros térmicos de solidificação.

Characterization of the Al-3wt.%Si alloy in unsteady-state horizontal directional solidification

The main purpose of this paper is to investigate both the columnar to equiaxed transition and primary dendritic arm spacings of Al-3wt.%Si alloy during the horizontal directional solidification. The transient heat transfer coefficient at the metal-mold interface is calculated based on comparisons between the experimental thermal profiles in castings and the simulations provided by a finite difference heat flow program. Simulated curve of the interfacial heat transfer coefficient was used in another numerical solidification model to determine theoretical values of tip growth rates, cooling rates and thermal gradients that are associated with both columnar to equiaxed transition and primary dendritic arm spacings. A good agreement was observed between the experimental values of these thermal variables and those numerically simulated for the alloy examined. A comparative analysis is carried out between the experimental data of this work and theoretical models from the literature that have been proposed to predict the primary dendritic spacings. In this context, this study may contribute to the understanding of how to manage solidification operational parameters aiming at designing the microstructure of Al-Si alloys.

Does radial growth of the lichen Parmelia conspersa depend exclusively on growth processes at the lobe tip?

The objective of this study was to test the hypothesis that the radial growth of lobes of the lichen Parmelia conspersa depends largely on growth processes which occur at the lobe tip. First, individual lobes were removed from thalli and portions of the lobe removed to within various distances from the tip. Radial growth of the lobe was unaffected until less than 2 mm of the lobe tip remained. Second, the surfaces of individual lobes were painted with acrylic paint leaving different portions of the lobe exposed. Painting lobes to within 0.5 mm and 1 mm of the tip substantially reduced radial growth. Third, the levels of ribitol, arabitol and mannitol were measured in different regions behind the lobe tip on four occasions during 1994. The concentration of the three carbohydrates was greatest at the lobe tip and the levels declined linearly with distance from the tip. Fourth, painting one vertical half of the lobe tip did not affect radial growth but artificially bisecting the lobe tip with a scalpel reduced radial growth. Although transport of carbohydrate from other regions of the lobe cannot be ruled out, the results support the hypothesis that radial growth in P. conspersa depends largely on processes within a region approximately 2 mm behind the lobe tip.

Fatigue behaviour of friction stir welded AA2024-T3 alloy: longitudinal and transverse crack growth

The fatigue crack growth properties of friction stir welded joints of 2024-T3 aluminium alloy have been studied under constant load amplitude (increasing-Delta K), with special emphasis on the residual stress (inverse weight function) effects on longitudinal and transverse crack growth rate predictions (Glinka`s method). In general, welded joints were more resistant to longitudinally growing fatigue cracks than the parent material at threshold Delta K values, when beneficial thermal residual stresses decelerated crack growth rate, while the opposite behaviour was observed next to K-C instability, basically due to monotonic fracture modes intercepting fatigue crack growth in weld microstructures. As a result, fatigue crack growth rate (FCGR) predictions were conservative at lower propagation rates and non-conservative for faster cracks. Regarding transverse cracks, intense compressive residual stresses rendered welded plates more fatigue resistant than neat parent plate. However, once the crack tip entered the more brittle weld region substantial acceleration of FCGR occurred due to operative monotonic tensile modes of fracture, leading to non-conservative crack growth rate predictions next to K-C instability. At threshold Delta K values non-conservative predictions values resulted from residual stress relaxation. Improvements on predicted FCGR values were strongly dependent on how the progressive plastic relaxation of the residual stress field was considered.

Inheritance of growth habit detected by genetic linkage analysis using microsatellites in the common bean (Phaseolus vulgaris L.)

The genetic linkage map for the common bean (Phaseolus vulgaris L.) is a valuable tool for breeding programs. Breeders provide new cultivars that meet the requirements of farmers and consumers, such as seed color, seed size, maturity, and growth habit. A genetic study was conducted to examine the genetics behind certain qualitative traits. Growth habit is usually described as a recessive trait inherited by a single gene, and there is no consensus about the position of the locus. The aim of this study was to develop a new genetic linkage map using genic and genomic microsatellite markers and three morphological traits: growth habit, flower color, and pod tip shape. A mapping population consisting of 380 recombinant F10 lines was generated from IAC-UNA x CAL143. A total of 871 microsatellites were screened for polymorphisms among the parents, and a linkage map was obtained with 198 mapped microsatellites. The total map length was 1865.9 cM, and the average distance between markers was 9.4 cM. Flower color and pod tip shape were mapped and segregated at Mendelian ratios, as expected. The segregation ratio and linkage data analyses indicated that the determinacy growth habit was inherited as two independent and dominant genes, and a genetic model is proposed for this trait.

Alterations in growth and branching of Neurospora crassa caused by sub-inhibitory concentrations of antifungal agents

Six antifungal agents at subinhibitory concentrations were used for investigating their ability to affect the growth and branching in Neurospora crassa. Among the antifungals herein used, the azole agent ketoconazole at 0.5 mu g/ml inhibited radial growth more than fluconazole at 5.0 mu g/ml while amphotericin B at 0.05 mu g/ml was more effective than nystatin at 0.05 mu g/ml. Morphological alterations in hyphae were observed in the presence of griseofulvin, ketoconazole and terbinafine at the established concentrations. The antifungal agents were more effective on vegetative growth than on conidial germination. Terbinafine markedly reduced growth unit length (GU) by 54.89%, and caused mycelia to become hyperbranched. In all cases, there was a high correlation between hyphal length and number of tips (r > 0.9). All our results showed highly significant differences by ANOVA, (p < 0.001, alpha = 0.05). Considering that the hyphal tip is the main interface between the fungus and its environment/through which enzymes and toxins are secreted and nutrients absorbed, it would not be desirable to obtain a hyperbranched mycelia with inefficient doses of antifungal drugs.

Modelling fungal growth on surfaces

Fungal growth in time and space at the substrate surface was modelled for a simple system mimicking solid-state fermentation, using a polycarbonate Nucleopore membrane laid over a glucose solution. Biomass production depends on both tip density and the diffusion of glucose within the fungal hyphae. The model predicts early increases in both height and concentration, followed by a period in which the biomass profile moves with a constant wavefront. The rate of increase in height increases as tip diffusivity increases or as the Monod saturation constant for glucose decreases.