Differential uptake, partitioning and transfer of Cd and Zn in the soil-pea plant-aphid system

The biomagnification of trace metals during transfer from contaminated soil to higher trophic levels may potentially result in the exposure of predatory arthropods to toxic concentrations of these elements. This study examined the transfer of Cd and Zn in a soil−plant−arthropod system grown in series of field plots that had received two annual applications of municipal biosolids with elevated levels of Cd and Zn. Results showed that biosolids amendment significantly increased the concentration of Cd in the soil and the shoots of pea plants and the concentration of Zn in the soil, pea roots, shoots, and pods. In addition, the ratio of Cd to Zn concentration showed that Zn was preferentially transferred compared to Cd through all parts of the system. As a consequence, Zn was biomagnified by the system whereas Cd was biominimized. Cd and Zn are considered to exhibit similar behaviors in biological systems. However, the Cd/Zn ratios demonstrated that in this system, Cd is much less labile in the root−shoot−pod and shoot−aphid pathways than Zn.

Perennial legumes native to Australia: a preliminary investigation of nutritive value and response to cutting

Six Australian native herbaceous perennial legumes (Lotus australis, Swainsona colutoides, Swainsona swainsonioides, Cullen tenax, Glycine tabacina and Kennedia prorepens) were assessed in the glasshouse for nutritive value, soluble condensed tannins and production of herbage in response to three cutting treatments (regrowth harvested every 4 and 6 weeks and plants left uncut for 12 weeks). The Mediterranean perennial legumes Medicago sativa and Lotus corniculatus were also included. Dry matter (DM) yield of some native legumes was comparable to L. corniculatus, but M. sativa produced more DM than all species except S. swainsonioides after 12 weeks of regrowth. Dry matter yield of all native legumes decreased with increased cutting frequency, indicating a susceptibility to frequent defoliation. Shoot in vitro dry matter digestibility (DMD) was high (>70%) in most native legumes, except G. tabacina (65%) and K. prorepens (55%). Crude protein ranged from 21-28% for all legumes except K. prorepens (12%). More frequent cutting resulted in higher DMD and crude protein in all species, except for the DMD of C. tenax and L. australis, which did not change. Concentrations of soluble condensed tannins were 2-9 g/kg DM in the Lotus spp., 10-18 g/kg DM in K. prorepens and negligible (<1 g/kg) in the other legumes. Of the native species, C. tenax, S. swainsonioides and L. australis showed the most promise for use as forage plants and further evaluation under field conditions is now warranted.

Root distributions of Australian herbaceous perennial legumes in response to phosphorus placement

Many Australian plant species have specific root adaptations for growth in phosphorus-impoverished soils, and are often sensitive to high external P concentrations. The growth responses of native Australian legumes in agricultural soils with elevated P availability in the surface horizons are unknown. The aim of these experiments was to test the hypothesis that increased P concentration in surface soil would reduce root proliferation at depth in native legumes. The effect of P placement on root distribution was assessed for two Australian legumes, Kennedia prorepens F. Muell. and Lotus australis Andrews, and the exotic Medicago sativa L. Three treatments were established in a low-P loam soil: amendment of 0.15 g mono-calcium phosphate in either (i) the top 50 mm (120 µg P g–1) or (ii) the top 500 mm (12 µg P g–1) of soil, and an unamended control. In the unamended soil M. sativa was shallow rooted, with 58% of the root length of in the top 50 mm. K. prorepens and L. australis had a more even distribution down the pot length, with only 4 and 22% of their roots in the 0–50 mm pot section, respectively. When exposed to amendment of P in the top 50 mm, root length in the top 50 mm increased 4-fold for K. prorepens and 10-fold for M. sativa, although the pattern of root distribution did not change for M. sativa. L. australis was relatively unresponsive to P additions and had an even distribution of roots down the pot. Shoot P concentrations differed according to species but not treatment (K. prorepens 2.1 mg g–1, L. australis 2.4 mg g–1, M. sativa 3.2 mg g–1). Total shoot P content was higher for K. prorepens than for the other species in all treatments. In a second experiment, mono-ester phosphatases were analysed from 1-mm slices of soil collected directly adjacent to the rhizosphere. All species exuded phosphatases into the rhizosphere, but addition of P to soil reduced phosphatase activity only for K. prorepens. Overall, high P concentration in the surface soil altered root distribution, but did not reduce root proliferation at depth. Furthermore, the Australian herbaceous perennial legumes had root distributions that enhanced P acquisition from low-P soils.

Contrasting behaviour of cadmium and zinc in a soil–plant–arthropod system

This study investigates the transfer of Cd and Zn from a soil amended with sewage sludge at rates up to 100 t ha(-1) through a multi-trophic system consisting of barley, the aphid Sitobion avenae and the larvae of the lacewing Chrysoperla carnae. Results show marked differences in the transfer of the two metals. Cadmium was freely accumulated in barley roots, but accumulation in the shoot was restricted to a concentration of around 0.22 mg kg(-1) (dry weight). This limited the transfer of Cd to higher trophic levels and resulted in no significant accumulation of Cd in S. avenae or in C. carnae. Zinc transfer in the system was largely unrestricted, resulting in significant accumulation in roots and shoots, in S. avenae and in C. carnae. Cadmium biomagnification occurred in lacewing pupae, with concentrations up to 3.6 times greater than in aphids. S. avenae biomagnified Zn by a factor of ca. 2.5 at low sludge amendment rates, but biomagnification decreased to a factor of 1.4 at the highest amendment rate. Biomagnification of Zn did not occur in C. carnae, but concentrations were up to 3.5 time higher than in soil. Results are discussed in light of the mechanisms regulating transfer of the two metals in the system.

Regurgitated bodies: presenting and representing trauma in 'The Act of Killing'

Focusing on The Act of Killing, this chapter examines how an “ethics of realism” operates on three key cinematic arenas: genre, authorship and spectatorship. As far as genre is concerned, the film’s realist commitment emerges from where it is least expected, namely from Hollywood genres, such as the musical, the film noir and the western, which are used as documentary, that is to say, as a fantasy realm where perpetrators can confess to their crimes without restraints or fear of punishment, but which nonetheless retains the evidentiary weight of the audiovisual medium. Authorship, in turn, translates as Oppenheimer’s unmistakable auteur signature through his role of self-confessed “infiltrator” who disguises as a sympathiser of the criminals in order to gain first-hand access to the full picture of their acts. One of them, the protagonist Anwar Congo, is clearly affected by post-traumatic stress disorder, and his repetitive reliving of his killings is made to flare up in front of the camera so as to bring back the dead to the present time in their material reality, through his own body, including a harrowing scene of the actor’s unpredictable and uncontrollable retching as he re-enacts the killing of his victims through strangulation. Finally, in the realm of spectatorship, the usual process of illusionistic identification on the part of the spectator is turned onto its head by means of disguising these criminals as amateur filmmakers, led to shoot, act within, and then watch their own film within the film so as to force them to experience beyond any illusion the suffering they had caused.

Analysis of phosphorus use efficiency traits in Coffea genotypes reveals Coffea arabica and Coffea canephora have contrasting phosphorus uptake and utilization efficiencies

Background and Aims: Phosphate (Pi) is one of the most limiting nutrients for agricultural production in Brazilian soils due to low soil Pi concentrations and rapid fixation of fertilizer Pi by adsorption to oxidic minerals and/or precipitation by iron and aluminum ions. The objectives of this study were to quantify phosphorus (P) uptake and use efficiency in cultivars of the species Coffea arabica L. and Coffea canephora L., and group them in terms of efficiency and response to Pi availability. Methods: Plants of 21 cultivars of C. arabica and four cultivars of C. canephora were grown under contrasting soil Pi availabilities. Biomass accumulation, tissue P concentration and accumulation and efficiency indices for P use were measured. Key Results: Coffee plant growth was significantly reduced under low Pi availability, and P concentration was higher in cultivars of C. canephora. The young leaves accumulated more P than any other tissue. The cultivars of C. canephora had a higher root/shoot ratio and were significantly more efficient in P uptake, while the cultivars of C. arabica were more efficient in P utilization. Agronomic P use efficiency varied among coffee cultivars and E16 Shoa, E22 Sidamo, Iêmen and Acaiá cultivars were classified as the most efficient and responsive to Pi supply. A positive correlation between P uptake efficiency and root to shoot ratio was observed across all cultivars at low Pi supply. These data identify Coffea genotypes better adapted to low soil Pi availabilities, and the traits that contribute to improved P uptake and use efficiency. These data could be used to select current genotypes with improved P uptake or utilization efficiencies for use on soils with low Pi availability and also provide potential breeding material and targets for breeding new cultivars better adapted to the low Pi status of Brazilian soils. This could ultimately reduce the use of Pi fertilizers in tropical soils, and contribute to more sustainable coffee production.

Propagation, growth, and carbohydrates of Dendrobium Second Love (Orchidaceae) in vitro as affected by sucrose, light, and dark

In general, plant material grown in vitro has low photosynthetic ability to achieve positive carbon balances. Therefore, a continuous supply of carbohydrates from the culture medium is required, and sucrose has been the most commonly used carbon source. In this paper, we investigate the effects of different sucrose concentrations and the presence and absence of light on the endogenous levels of soluble carbohydrates and starch as well as on the proliferation and growth of Dendrobium Second Love (Orchidaceae) in vitro. The possibility of using etiolated stem segments as a means for micropropagating this hybrid was also verified. The results obtained indicated that the presence and absence of light and the sucrose concentrations used influenced the amounts of soluble carbohydrates and starch and the proliferation of D. Second Love shoots and roots. An increase in sucrose concentration caused a progressive increase in the amounts of total carbohydrates and starch. Under both light conditions, sucrose was the main sugar found in the shoots followed by glucose and fructose. The addition of sucrose to the culture medium up to 2% and 4% was advantageous to the number of shoots produced per explant and the root longitudinal growth in the presence and absence of light, respectively. Shoot and root dry matter and the number of roots formed per explant increased as sucrose concentration was raised up to 6% in both light treatments. The use of dark-grown shoot segments proved to be a useful and reliable alternative for the micropropagation of this hybrid.

Ethylene and polyamine interactions in morphogenesis of Passiflora cincinnata: effects of ethylene biosynthesis and action modulators, as well as ethylene scavengers

Ethylene is a plant hormone that is of fundamental importance to in vitro morphogenesis, but in many species, it has not been thoroughly studied. Its relationship with polyamines has been studied mainly because the two classes of hormones share a common biosynthetic precursor, S-adenosylmethionine (SAM). In order to clarify whether competition between polyamines and ethylene influences in vitro morphogenetic responses of Passiflora cincinnata Mast., a climacteric species, different compounds were used that act on ethylene biosynthesis and action, or as ethylene scavengers. Treatment with the ethylene inhibitor, aminoethoxyvinylglycine (AVG) caused a greater regeneration frequency in P. cincinnata, whereas treatment with the ethylene precursor, 1-aminocyclopropane-1-carboxylic-acid (ACC) lessened regeneration frequencies. The data suggested that levels of polyamines and ethylene are not correlated with morphogenic responses in P. cincinnata. It was ascertained that neither the absolute ethylene and polyamine levels, nor competition between the compounds, correlated to the obtained morphogenic responses. However, sensitivity to, and signaling by, ethylene appears to play an important role in differentiation. This study reinforces previous reports regarding the requirement of critical concentrations and temporal regulation of ethylene levels for morphogenic responses. Temporal regulation also appeared to be a key factor in competition between the two biosynthetic pathways, without having any effects on morphogenesis. Further studies investigating the silencing or overexpression of genes related to ethylene perception, under the influence of polyamines in cell differentiation are extremely important for the complete understanding of this process.

Expression pattern of four storage xyloglucan mobilization-related genes during seedling development of the rain forest tree Hymenaea courbaril L.

During seedling establishment, cotyledons of the rain forest tree Hymenaea courbaril mobilize storage cell wall xyloglucan to sustain growth. The polysaccharide is degraded and its products are transported to growing sink tissues. Auxin from the shoot controls the level of xyloglucan hydrolytic enzymes. It is not yet known how important the expression of these genes is for the control of storage xyloglucan degradation. In this work, partial cDNAs of the genes xyloglucan transglycosylase hydrolase (HcXTH1) and beta-galactosidase (HcBGAL1), both related to xyloglucan degradation, and two other genes related to sucrose metabolism [alkaline invertase (HcAlkIN1) and sucrose synthase (HcSUS1)], were isolated. The partial sequences were characterized by comparison with sequences available in the literature, and phylogenetic trees were assembled. Gene expression was evaluated at intervals of 6 h during 24 h in cotyledons, hypocotyl, roots, and leaves, using 45-d-old plantlets. HcXTH1 and HcBGAL1 were correlated to xyloglucan degradation and responded to auxin and light, being down-regulated when transport of auxin was prevented by N-1-naphthylphthalamic acid (NPA) and stimulated by constant light. Genes related to sucrose metabolism, HcAlkIN1 and HcSUS1, responded to inhibition of auxin transport in consonance with storage mobilization in the cotyledons. A model is proposed suggesting that auxin and light are involved in the control of the expression of genes related to storage xyloglucan mobilization in seedlings of H. courbaril. It is concluded that gene expression plays a role in the control of the intercommunication system of the source-sink relationship during seeding growth, favouring its establishment in the shaded environment of the rain forest understorey.

Cearania heterophylla gen. nov et sp nov., a fossil gymnosperm with affinities to the Gnetales from the Early Cretaceous of northern Gondwana

Vegetative and fertile shoots of a shrub-like seed plant from the late Aptian Crato Formation of Brazil are described as Cearania heterophylla Kunzmann, Mohr and Bernardes-de-Oliveira, gen. nov. et sp. nov. Anatomical details of the axes, epidermal features and separate ovulate and pollen producing organs indicate the gymnospermous nature of this plant. The vascular tissue of the axes includes tracheids with bordered pits and fiber tracheids. Vegetative shoots comprising at least three branching orders bear opposite-decussately arranged ovate to lanceolate, dorsiventrally flattened, parallelodromous, rather thick leaves that vary tremendously in size. The amphistomatic leaves bear (brachy-)paracytic stomatal complexes arranged in simple longitudinal files. The ovulate structure is interpreted as a terminally attached single globular ovule/seed surrounded by at least five to six lanceolate bracts. A terminally attached pollen-cone like structure grows on a lateral leafy shoot. The unusual character combination may indicate that the fossils belong to a hitherto unknown group with affinities to ephedroid Gnetales. Sterile shoots formerly often described as Podozamites, Nageiopsis or Lilites that are at least partly congeneric with C. heterophylla Kunzmann, Mohr and Bernardes-de-Oliveira, gen. nov. et sp. nov. had a wide geographic distribution during the Early Cretaceous. (C) 2009 Elsevier B.V. All rights reserved.

Molecular characterization of Arabidopsis thaliana PUF proteins - binding specificity and target candidates

PUF proteins regulate both stability and translation through sequence-specific binding to the 3` UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3` UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3` UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants.

INSECT CHYMOTRYPSINS: CHLOROMETHYL KETONE INACTIVATION AND SUBSTRATE SPECIFICITY RELATIVE TO POSSIBLE COEVOLUTIONAL ADAPTATION OF INSECTS AND PLANTS

Insect digestive chymotrypsins are present in a large variety of insect orders but their substrate specificity still remains unclear. Ewer insect chymotrypsins from 3 different insect orders (Dictyoptera, Coleoptera and two Lepidoptera) were isolated using affinity chromatography. Enzymes presented molecular masses in the range of 20 to 31 kDa and pH optima in the range of 7.5 to 10.0. Kinetic characterization. using different, colorimetric and fluorescent substrates indicated that insect chymotrypsins differ from, bovine chymotrypsin in their primary specificity toward small substrates (like N-benzoyl-L-Tyr p-nitroanilide) rather than on their preference for large substrates (exemplified by Succynil-Ala-Ala-Pro-Phe P-nitroanilide). Chloromethyl ketones (TPCK, N-alpha-tosyl-L-Phe chloromethyl ketone and Z-GGF-CK, N-carbobenzoxy-Gly-Gly-phe-CK) inactivated all chymotrypsins legated. Inactivation rates follow apparent first-order kinetics with variable second order rates (TPCK, 42 to 130 M(-1)s(-1); Z-GGF-CK, 150 to 450 M(-1)s(-1) that may be remarkably low for S. frugiperda chymotrypsin (TPCK, 6 M(-1)s(-1); Z-GGF-CK, 6.1 M(-1) s(-1)). Homology modelling and sequence alignment showed that. in lepidopteran chymotrypsins, differences in the amino acid residues in the neighborhood of the catalytic His 57 may affect its pKa, value. This is Proposed as the cause of the decrease in His 57 reactivity toward chloromethyl ketones. Such amino acid replacement in the active site is proposed. to be an adaptation to the presence of dietary ketones. (C) 2009 Wiley Periodicals, Inc.

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)

In vitro morphogenesis and cell suspension culture establishment in Piper solmsianum C. DC. (Piperaceae)). Piper solmsianum is a shrub from Southeast Brazil in which many biologically active compounds were identified. The aim of this work was to establish a cell suspension culture system for this species. With this in mind, petiole and leaf explants obtained from in vitro plantlets were cultured in the presence of different plant growth regulator combinations (IAA, NAA, 2,4-D and BA). Root and indirect shoot adventitious formation, detected by histological analysis, was observed. Besides the different combinations of plant growth regulators, light regime and the supplement of activated charcoal (1.5 mg.l(-1)) were tested for callus induction and growth. Cultures maintained in light, on a 0.2 mg.l(-1) 2,4-D and 2 mg.l(-1) BA supplemented medium, and in the absence of activated charcoal, showed the highest calli fresh matter increment. From a callus culture, cell suspension cultures were established and their growth and metabolite accumulation studied. The achieved results may be useful for further characterization of the activated secondary metabolites pathways in in vitro systems of P. solmsianum.

Cultura de anteras e embriogênese de genótipos selecionados de cevada (Hordeum vulgare L.ssp.vulgare)

A eficiência da técnica de cultura de anteras, em escala comercial, ainda pode ser considerada baixa quando medida em número de plantas duplo-haplóides férteis obtidas para cada antera estabelecida in vitro. Dessa forma, o presente trabalho é pioneiro no estudo detalhado da embriogênese in vitro do micrósporo e do grão de pólen de cevada (Hordeum vulgare L. ssp. vulgare). Com o objetivo de contribuir para o aperfeiçoamento da técnica de cultura de anteras foi analisada a embriogênese, com especial ênfase na etapa da indução, através de análises citológicas e histológicas de anteras cultivadas in vitro. Foram analisadas uma cultivar brasileira de cevada, em comparação com linhagens de duas outras cultivares brasileiras, que foram selecionadas, por seleção divergente para maior ou para menor resposta na indução da rota embriogênica e, respectivamente, para menor ou para maior capacidade de regenerar plântulas verdes. Somente foram estabelecidas em cultivo in vitro as anteras que apresentaram micrósporos e pólens jovens, das linhagens selecionadas da cultivar A-05 (S3A22 e S3A23), e da cultivar BR-2(S3B63 e, apenas na cultura de anteras, S3B61), bem como da cultivar MN-599 (nãoselecionada). Para as análises histológicas, foram fixadas, a cada dois dias, duas anteras, correspondentes a cada fileira da mesma espiga, após o início do cultivo in vitro. As anteras em cultivo e respectivas estruturas multicelulares foram fixadas em FAA 50%, desidratadas em série etílica e incluídas em hidroxietilmetacrilato. Os blocos de resina polimerizada foram secionados longitudinalmente com 3 mm de espessura. Para as análises citológicas foram fixadas, de cada espiga recém-coletada, três espiguetas sendo uma da base, outra do meio e outra do ápice. Após o pré-tratamento à baixa temperatura (5 °C), porém antes do cultivo in vitro, foram fixadas três anteras (amostras utilizadas como controles). A cada três dias, durante o cultivo, três anteras foram fixadas (até 18 dias). As anteras em cultivo e estruturas multicelulares foram fixadas em Farmer e FAA 50%, transferidas após 24 horas para etanol 70%. Na cultura in vitro das anteras houve diferenças entre uma das linhagens da cultivar A-05 em relação a cultivar MN- 599, na produção inicial de estruturas embriogênicas, diferença que desapareceu na produção total. Entretanto, houve diferenças na formação dos xiii embriões: a cv.MN-599 formou embriões bem diferenciados ao passo que a linhagem S3A22 produziu um número aparentemente menor, sendo que os embriões não eram bem diferenciados. A linhagem S3B63 não apresentou embriões até o final da análise histológica. Considerando que a amostra dessa linhagem, mantida em cultura, formou plantas verdes, pode-se propor que a formação de embriões deve ocorrer posteriormente ao desenvolvimento da cv.MN-599. Cabe destacar que houve diferenças significativas entre as cultivares A-05 e BR-2 quanto à regeneração de plântulas verdes. Esses resultados indicam ter havido maior eficiência da seleção em relação à etapa da regeneração. Com relação às categorias classificatórias dos micrósporos e grãos de pólen, constatou-se que desde o início da análise histológica (2o dia de cultivo in vitro) até o final (34o dia), foram observados micrósporos, o mesmo tendo sido observado na análise citológica. Os grãos de pólen multinucleados ocorreram praticamente em todo o período de cultivo in vitro, em ambas análises; não ocorrendo nos controles da citologia (antes do cultivo); os multinucleados foram observados a partir do 3o dia, enquanto que os multicelulares a partir do 4o dia de cultivo. As estruturas multicelulares foram observadas a partir do 8o dia. A quantidade e o tamanho das estruturas multicelulares foram variáveis ao longo da análise histológica, sendo que do 14o ao 20o dia foram encontradas as de maiores dimensões, resultantes da proliferação celular por mitoses sucessivas. A partir do 22o dia (cultivar MN- 599), a ocorrência de estruturas multicelulares no interior dos lóculos da antera diminuiu, predominando o processo de proliferação externo às anteras. Para as linhagens, a partir do 18o dia foram observadas estruturas multicelulares liberadas das anteras. A análise das estruturas multicelulares permitiu classificá-las em quatro categorias: 1. SFD: Sem forma definida; 2. MAC: meristema apical caulinar; 3. MAR: meristema apical radical embrionário adventício; e 4. Embriões. As estruturas amorfas apareceram em maior número, quando comparadas com as outras categorias. Em síntese: as linhagens selecionadas e a cultivar diferiram não apenas no tempo necessário para a formação dos embriões, mas também no desenvolvimento dos mesmos, que foi mais diferenciado na cultivar MN-599, porém sendo observados mais cedo na linhagem S3A22 e S3A23, do que na cultivar MN-599.

Desafios do modelo de franquias no setor de perfumaria e cosméticos brasileiro, sob o ponto de vista de franqueados e franqueadores

O objetivo fundamental deste trabalho foi investigar e entender quais são os desafios para que o modelo de franquias possa crescer ainda mais e adquirir maior representatividade enquanto modelo de distribuição no setor de perfumaria e cosméticos brasileiro. Para isto, utilizou-se, além de uma revisão bibliográfica a partir do referencial teórico disponível, uma abordagem qualitativa, segundo a qual foram realizadas vinte e duas entrevistas com franqueados e franqueadores, no período compreendido entre setembro e novembro de 2010. As empresas representadas por estes entrevistados foram: Água de Cheiro, Antídoto, Contém 1g, Mahogany e O Boticário. A realização destas entrevistas revelou que, embora o modelo de franquias seja visto por franqueados e franqueadores como uma boa alternativa para a distribuição de produtos e para o aumento da capilaridade da marca, o modelo de franquias apresenta alguns desafios a serem alcançados, tais como maior participação ativa da rede franqueadora, melhoria no modelo de avaliação e seleção de franqueados, melhor suporte financeiro aos franqueados nos primeiros anos após a abertura da loja, melhor modelo para escolha da localização das lojas e, por fim, melhor modelo para definição de taxas de royalties.