BACKGROUND:Sugarcane breeding has significantly progressed in the last 30 years, but achieving additional yield gains has been difficult because of the constraints imposed by the complex ploidy of this crop. Sugarcane cultivars are interspecific hybrids between Saccharum officinarum and Saccharum spontaneum. S. officinarum is an octoploid with 2n=80 chromosomes while S. spontaneum has 2n=40 to 128 chromosomes and ploidy varying from 5 to 16. The hybrid genome is composed of 70-80%S. officinaram and 5-20%S. spontaneum chromosomes and a small proportion of recombinants. Sequencing the genome of this complex crop may help identify useful genes, either per se or through comparative genomics using closely related grasses. The construction and sequencing of a bacterial artificial chromosome (BAC) library of an elite commercial variety of sugarcane could help assembly the sugarcane genome.RESULTS:A BAC library designated SS_SBa was constructed with DNA isolated from the commercial sugarcane variety SP80-3280. The library contains 36,864 clones with an average insert size of 125 Kb, 88% of which has inserts larger than 90 Kb. Based on the estimated genome size of 760-930 Mb, the library exhibits 5-6 times coverage the monoploid sugarcane genome. Bidirectional BAC end sequencing (BESs) from a random sample of 192 BAC clones sampled genes and repetitive elements of the sugarcane genome. Forty-five per cent of the total BES nucleotides represents repetitive elements, 83% of which belonging to LTR retrotransposons. Alignment of BESs corresponding to 42 BACs to the genome sequence of the 10 sorghum chromosomes revealed regions of microsynteny, with expansions and contractions of sorghum genome regions relative to the sugarcane BAC clones. In general, the sampled sorghum genome regions presented an average 29% expansion in relation to the sugarcane syntenic BACs.CONCLUSION:The SS_SBa BAC library represents a new resource for sugarcane genome sequencing. An analysis of insert size, genome coverage and orthologous alignment with the sorghum genome revealed that the library presents whole genome coverage. The comparison of syntenic regions of the sorghum genome to 42 SS_SBa BES pairs revealed that the sorghum genome is expanded in relation to the sugarcane genome.

Thesis (MSc)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: The changes in carbon allocation to sucrose, hexoses, fibre, starch and respiration were investigated in developing internodes of sugarcane. Radiolabelling studies were conducted on internode 3, 6 and 9 tissue, representing three stages of increasing maturity. It was apparent that a high rate of cycling between triose-phosphate and hexose-phosphate occurred. A maximum of 50% of carbon entering triose-phosphates was returned to hexose-phosphate in internode 3 tissue, and this flux decreased with tissue maturity to 30%. Carbon partitioning into sucrose increased from 34% of total 14C uptake in internode 3, to 61% in internodes 6 and 9. In immature tissue, the protein and fibre components were the dominant competing sinks with sucrose for incoming carbon, to which 14 and 16% of carbon were allocated respectively. Increased carbon allocation to sucrose with tissue maturity, coincided with a decrease in partitioning to fibre. This indicated that previous studies had underestimated total carbon allocation to respiration, since the protein component was not considered. In contrast with earlier work, the respiratory pathway was the strongest competitor with sucrose for incoming carbon, even in mature tissue. Between internodes 3 and 6, carbon allocation to total respiration did not change significantly, but decreased 50% in mature tissue. Starch was a weak competitor with sucrose, for incoming carbon, to which a maximum of 2% of 14Cwas allocated in immature tissue. In cane harvested in early spring, radiolabelled maltose was recovered in internode 3 tissue of ripening cane, indicating that concomitant starch synthesis and degradation occurred. The. redistribution of C-1 and C-6 in starch glucose was analysed following feeding of tissue with [1_14C]_and [6_14C]_glucose. Randomization of label in starch indicated that the pathway for carbon movement into sugarcane plastids for starch synthesis is primarily through the triose-phosphate translocator. Finally, this study indicated that radiolabelling of tissue discs is a suitable experimental system to determine carbon flux in sugarcane. During the 3 h labelling period the rate of 14C02 release became linear, indicating that the system approached isotopic steady state between the external and internal glucose pool; and the respiratory processes involved in CO2 release. / AFRIKAANSE OPSOMMING: Die veranderinge in koolstoftoedeling na sukrose, heksoses, vesel, stysel en respirasie is in ontwikkelende internodes van suikerriet ondersoek. Die koolhidraatmetabolisme in internodes 3, 6 en 9, wat drie stadiums van toenemende rypheid verteenwoordig, is met behulp van 14Cmerkingstudies ondersoek. Dit is duidelik dat daar 'n hoë mate van koolstofsirkulering tussen die heksose- en triose-fosfaat poele voorkom. In internode 3 word tot 50% van die koolstofwat in triose-fosfate geïnkorporeer is, weer na heksosefosfaat omgeskakel. Selfs in volwasse weefsel vind daar nog soveel as 30% koolstofsirkukering plaas tussen die twee poele plaas. Koolstoftoedeling vanaf glukose na sukrose het van 34% in internode 3, tot 61% in internodes 6 en 9 toegeneem. Proteïn en selwandkomponente was die belangrikste swelgpunte vir koolstof in onvolwasse weefsel gewees. Namate die weefsel meer volwasse word, word sukrose 'n belangriker swelgpunt. Die koolstoftoedeling aan sukrose is veral ten koste van toedeling aan die selwandkomponente. Die bevinding dat die proteïenpoel 'n sterk swelgpunt is dui aan dat vorige studies die belang van respiratoriese koolstofvloei onderskat het. In teenstelling met vorige aansprake is dit duidelik dat selfs in volwasse weefsel respirasie die grootste swelpunt vir die inkomende organiese koolstof in die internode vorm. Koolstoftoedeling aan respirasie het nie noemenswaardig tussen internodes 3 en 6 verskil nie, maar het met 50% in volwasse weefsel afgeneem. Stysel is deurgaans 'n swak swelgpunt vir koolstof met die hoogste toedeling aan die poel (2%) in die jong weefsel (internode 3) . Na toediening van [U- 14C]-glukose is radioaktief gemerkte maltose gevind in suikerriet wat vroeg in die lente geoes is. Dit dui aan dat gelyktydige afbraak en sintese van stysel plaasgevind het. Die herverdeling van C-l en C-6 in glukose afkomstig van stysel is na toediening van [1_14C]_ en [6-14C]-glukose ontleed. Die ewekansige verspreiding van radioaktiwiteit tussen koolstof 1 en 6 van die glukose in stysel dui aan dat dit hoofsaaklik die triose-fosfaat translokeerder is wat in die plastied verantwoordelik is. Hierdie studie het ook aangetoon dat radioaktiewe merking van weefselsnitte 'n geskikte eksperimentele sisteem is om koolstoftoedeling in suikerriet te ondersoek. Die patroon van 14C02 vrystelling dui daarop dat die weefsel na 'n 3 h inkuberingsperiode isotopiese ewewig tussen die eksterne en interne glukose poele en die respiratoriese prosesse begin bereik het.

Dissertation (PhD)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: This study had three main goals: 1. to investigate the occurrence on the protein level of sucrose synthase (SuSy) isoforms in sugarcane sink tissue, 2. to determine the kinetic properties of these isoforms, 3. to establish the tissue localisation of SuSy in the sugarcane culm The results are summarised below: Three SuSy isoforms were obtained from leaf roll tissue. The SuSyA and SuSyB isoforms differed in terms of charge characteristics, with SuSyA not binding to an anion exchange column that bound SuSyB and SuSyC under the same conditions. Both SuSyB and SuSyC isoforms were eluted at 180 mM KCl. The SuSyA and SuSyB isoforms were present during autumn, but during winter only the SuSyC isoform could be isolated. Even though they eluted at the same salt concentration, SuSyB and SuSyC were different isoforms, because they had different kinetic parameters, as well as different immunological properties. SuSyB and SuSyC could not have been mixtures of the same isoforms, since a polyclonal antiserum against SuSyB, which inactivates native SuSyB, did not inactivate SuSyC. All three isoforms had significantly different kinetic parameters, with the SuSyA isoform also having a much lower sucrose breakdown/synthesis ratio than the other two isoforms. Therefore, at least three SuSy isoforms occur in sugarcane leaf roll tissue on the protein level. The SuSyC isoform was subsequently kinetically characterised in detail. Data showed that the enzyme employs an ordered ternary complex mechanism, with UDP binding first and UDP-glucose dissociating last. These experimentally obtained kinetic parameters were then used to extend a kinetic model of sucrose accumulation. Data show that when the experimentally determined SuSy kineticparameters were entered into the model, a 40 % increase in sucrose concentration and 7 times reduction in fructose concentration resulted. These data illustrate the pronounced physiological effects that may result from the presence of different SuSy isoforms. SuSy protein localisation data, obtained by an immunohistochemical approach, indicated that SuSy protein was present in both storage parenchyma and vascular tissue of young, intermediate, and mature internodes. SuSy enzyme activity in different parts of the internodes was similar, except for internode 3, which had much higher activity in the bottom part of the internode, possibly because growth is faster here, hence a higher demand for sucrose cleavage exists here. / AFRIKAANSE OPSOMMING: Hierdie studie het ten doel gehad: 1. om die teenwoordigheid van sukrose sintase (SuSy) isovorme in suikkerriet swelgweefsel te ondersoek 2. om die kinetiese eienskappe van hierdie isovorme te ondersoek 3. om die weefsellokalisering van SuSy in die suikerrietstingel te bepaal Die resultate word hieronder opgesom: Drie SuSy isovorme is gevind in blaarrol weefsel. Die SuSyA en SuSyB isovorme het verskil in terme van ladingseienskappe, met SuSyA wat nie aan ‘n anioonuitruilkolom gebind het nie waaraan SuSyB en SuSyC wel onder dieselfde kondisies gebind het. Beide SuSyB en SuSyC isovorme is geëlueer van die kolom teen 180 mM KCl. Die SuSyA en SuSyB isovorme was teenwoordig gedurende herfs, maar in die winter was slegs SuSyC teenwoordig. Ten spyte van die feit dat SuSyB en SuSyC teen dieselfde soutkonsentrasie geëlueer is, het hulle verskillende isovorme verteenwoordig, aangesien hulle kinetiese en immunologiese eienskappe verskil het. SuSyB en SuSyC kon nie mengsels van dieselfde isovorme gewees het nie, want ‘n poliklonale antiserum teen SuSyB, wat SuSyB geïnaktiveer het, het nie SuSyC geïnaktiveer nie. Al drie isovorme het betekenisvol verskil wat kinetiese eienskappe betref, met die SuSyA isovorm wat ook ‘n baie laer sukrose afbraak/sintese verhouding gehad het as die ander twee isovorme. Daar is dus ten minste drie SuSy isovorme teenwoordig op die proteïen vlak in suikerriet blaarrol weefsel. Die in-detail kinetiese analise van die SuSyC isovorm het getoon dat die ensiem ‘n geordende drietallige kompleks meganisme het, met UDP wat eerste bind en UDP-glukose wat laaste dissosieer. Die eksperimenteel bepaalde kinetiese parameters is toe gebruik om ‘n kinetiese model van sukrose akkumulering uit tebrei. Data het getoon dat wanneer die generiese SuSy kinetiese parameters in die oorspronklike model vervang word met die eksperimenteel bepaalde waardes, die berekende sukrose konsentrasie met ongeveer 40 % toeneem, terwyl die fruktose konsentrasie ongeveer 7 keer afneem. Hierdie resultaat toon die groot fisiologiese effek wat die uitdrukking van verskillende SuSy isovorme op suikermetabolisme kan hê. Die SuSy proteïen lokaliseringsdata, wat met ‘n immunohistochemiese benadering verkry is, het aangedui dat SuSy in beide bergingsparenchiemselle sowel as vaatweefsel teenwoordig is in jong, intermediêre en volwasse internodes. SuSy ensiemaktiwiteit in verskillende dele van die internodes was soortgelyk, behalwe in internode 3, wat baie hoër aktiwiteit gehad het in die onderste deel van die internode as bo, moontlik weens vinniger groei in hierdie deel van die internode, wat afhanklik is van afbraakprodukte van sukrose.

Chromatographic methods were used to determine the absolute values of cis-aconitic acid and trans-aconitic acid present in sugar cane factory processing streams and to determine the rates of isomerisation from the trans-aconitic acid to the cis-aconitic acid isomer. A reproducible, solid phase, ion exchange extraction method was developed to isolate the organic acids found in sugar factory process streams. The isolated acids were quantified using a dual ion-exclusion column, high performance liquid chromatographic method. To improve resolution of the acids, columns were maintained at different temperatures, whilst the combined use of both ultra-violet (UV) and refractive index detection proved useful in peak identification. Concentrations of the aconitic acid isomers were used to calculate the cis/trans aconitic acid isomer ratio occurring across the different processes found in a sugar cane factory. trans-Aconitic acid was found to be the predominant form present in the cane entering the factory. Analysis showed that isomerisation of the trans-aconitic acid to the cis-aconitic acid isomer occurred during processing. To understand and model this reaction, a reproducible experimental isomerisation method was developed making use of buffers to maintain pH conditions during experiments. A chromatographic analysis method, using ion-exclusion chromatography and UV detection, was developed to analyse the isomerisation reaction mixture. Chromatography was used in both an on-line and off-line mode for quantitation of the isomers. The method was used to study the isomerisation under conditions similar to those found in the factory. These included pH, temperature, ionic strength and the presence of monovalent and divalent cations found in sugar cane juices. It was shown that the isomerisation is a first order reversible reaction under the conditions studied. Temperature and pH were shown to be the important isomerisation variables. Temperature enhances the rate of isomerisation of the trans-aconitic isomer to the cis-aconitic acid isomer whilst pH affects the ultimate cis/trans aconitic acid ratio attained. Ionic strength was found to be a relatively unimportant factor. The presence of divalent and monovalent cations, at concentrations usually found in cane juice, was shown to have little effect on the rate of isomerisation. Activation parameters, including the activation energy (Ea), pre-exponential factor (log A), enthalpy (?H‡) and entropy of activation (?S‡), were calculated at each combination of buffer concentration and pH used in the experimental procedures. The values recorded are of a similar value to those reported for structurally similar compounds. cis-Aconitic acid was shown to undergo decarboxylation to itaconic acid. This occurred at low pH values and high temperatures. A detailed study was not undertaken since the conditions under which it occurs are considered extreme from the viewpoint of a sugar technologist. A model describing the equilibrium cis/trans aconitic acid isomer ratio was developed as a function of pH, temperature and time from the kinetic results. This was used to predict the equilibrium ratio for the aconitic acid isomers at the output of various processes in the sugar factory. Given the time, average pH and temperature the model can successfully predict the equilibrium ratio for the relevant process stream. / Thesis (M.Sc.)-University of Natal, Durban, 2000.

O emprego de mudas pré-brotadas (MPB) de cana-de-açúcar, no sistema canavieiro, cresceu imensamente nos últimos anos. Entretanto, a carência de informações no manejo com herbicidas no sistema pode comprometer a tecnologia. Com o objetivo de compreender melhor os efeitos causados por diferentes herbicidas aplicados em pré e pós-transplantes das MPBs de cana-de-açúcar da variedade CTC4, este estudo foi conduzido em duas etapas em diferentes condições. Os tratamentos foram definidos com base nos herbicidas mais usuais do setor. Ambos os experimentos foram conduzidos no centro de pesquisa da FMC em Paulínia (SP), onde efeitos como fitotoxicidade, altura, produtividade, número de perfilhos, número de colmos e germinação foram avaliados. Para o experimento de campo, observou-se que apesar de injúrias iniciais causadas por diversos herbicidas, apenas o sulfentrazone promoveu redução de produtividade, entretanto todos tratamentos aos 60 dias após à aplicação não apresentavam sintomas de fitotoxidez. A partir desse primeiro estudo, foi realizado em casa de vegetação outro experimento, buscando analisar os mesmos herbicidas em diferentes posicionamentos de aplicação e em dois tipos de solo. Quando posicionado em pré transplante os sintomas de injúrias foram mais elevados, comprometendo o desenvolvimento de alguns tratamentos. Neste caso, o diclosulam mostrou-se o menos seletivo independentemente do tipo de solo estudado, seguido pelo clomazone. Entretanto, a literatura mostra que resultados de seletividade variam de acordo com as condições empregadas e principalmente variedades da cultura, assim, repetições deste estudo devem ser realizadas para possibilitar uma melhor compreensão e validação dos dados. / The use of pre-sprouted seedlings (PSS) of sugarcane in the sugarcane\'s system has grown immensely in recent years. However, lack of information on herbicide management in the system may compromise the technology. In order to better understand the effects caused by different herbicides applied in pre and post-transplantation of sugarcane MPBs variety CTC4, this study was conducted in two stages under different conditions. The treatments were defined based on the herbicides most common in the industry. Both experiments were conducted at the FMC research center in Paulínia (SP), where effects such as phytotoxicity, height, yield, number of tillers, number of stems and germination were evaluated. For the field experiment, it was observed that in spite of initial insults caused by several herbicides, only sulfentrazone promoted a reduction of productivity, however all treatments at 60 days after application did not present phytotoxicity symptoms. From this first study, another experiment was carried out in a greenhouse, trying to analyze the same herbicides in different application positions and in two types of soil. When positioned in pretransplantation the symptoms of injuries were higher, compromising the development of some treatment. In this case, diclosulam showed to be the least selective independent of the type of soil studied, followed by clomazone. However, the literature shows that selectivity results vary according to the conditions employed and mainly the crop varieties, thus, repetitions of this study should be performed to enable better understanding and validation of the data.

A Thesis, submitted to the Faculty of Science, University of the Witwatersrand, in fulfilment of the requirements for the degree of Doctor of Philosophy. Johannesburg, 2013 / A novel holistic approach was used to study the mycotrophic nature of commercial sugarcane varieties grown in KwaZulu-Natal, South Africa. All five varieties were mycotrophic, but N12 had the highest overall mycorrhization and was selected for a pot study to assess the growth response of sugarcane to inoculation with indigenous arbuscular mycorrhizal (AM) fungi and microflora. The pot study suggested that sugarcane will respond positively to inoculation with AM fungi, but the effects are most clear in the early phase of growth and less obvious in later elongation phases. This observation, taken together with the ability of sugarcane to grow well in sterile soil without microflora additions suggests that the plant may be facultatively mycotrophic. A multivariate analysis determined the nutrient relationships between soil and corresponding leaf nutrient levels on 72 sugarcane field plants, categorised according to either high or low percentage colonisation. Highly colonised plants were found to have more positive nutrient correlations compared to lower percentage colonised plants. AM fungi were identified from spore morphology and associated mycorrhizal bacteria (AMB) were identified by 16s rDNA analysis. Partial molecular identification was conducted using a universal eukaryotic forward NS31 primer and general fungal AM1 primers confirming the spores to be of AM fungi origin. A nested PCR was performed, using the universal fungal primers, NS5 and ITS4, followed by primer combinations to target sequences of specific Glomalean groups. Only partial molecular identification was conducted, as RFLPs were not successfully optimised. DNA from the Acaulospora gerdemannii/Acaulospora trappei group, Glomus occultum/Glomus brasilianum group, Glomus mosseae/Glomus intraradices group, Glomus etunicatum/ Glomus clariodeum group and Acaulosporaceae sensu stricto were detected, indicating AM fungi diversity. Bacteria, Brevibacillus reuszeri isolated from Scutellospora nigra, Bacillus megaterium and Stenotrophomonas maltophilia isolated from Glomus geosporum, Paenibacillus chitinolyticus and Bacillus cereus isolated from Acaulospora mellea and Gigaspora margarita spores respectively, were tested for biocontrol capability against pathogenic nematodes of Paratrichodorus, Meloidogyne and Pratylenchus genera. Meloidogyne was the least susceptible to associated mycorrhizal bacteria biocontrol and Paratrichodorus the most susceptible. These studies have contributed to understanding the role of AM in sugarcane agriculture in South Africa. Keywords: arbuscular mycorrhiza, sugarcane varieties, multivariate analysis, nutrient relationships, associated mycorrhizal bacteria, biocontrol, nematodes

Resumo: Avaliou-se a eficiência de surfactantes e espalhantes adesionantes na homogeneização da calda de pulverização, constituída por óleo fúsel e água. Para o teste, utilizou-se de Energic, detergente neutro, álcool e Haiten. Para erradicação da cana-de-açúcar comparou-se o óleo fúsel isolado e em mistura com glifosato, em vasos de 22 L, em delineamento inteiramente casualizado, com 12 tratamentos em 4 repetições. Em outro experimento, o óleo fúsel isolado foi aplicado em plantas daninhas em pré-plantio incorporado, em delineamento inteiramente casualizado, com 25 tratamentos em quatro repetições, analisado em esquema fatorial 5x5 (concentrações e espécies); em pós-emergência das plantas daninhas houve a mistura do óleo fúsel e glifosato, em diferentes concentrações. Por último, avaliou-se a eficiência do óleo fúsel isolado e em mistura com glifosato, aplicados em pós-emergência tardia de plantas daninhas, em delineamento em blocos casualizados com 13 tratamentos em 4 repetições. Na mistura do óleo à calda de pulverização, Energic e detergente neutro proporcionaram melhores resultados, enquanto álcool e Haiten mostraram-se ineficientes. O óleo fúsel isolado não dessecou a cana-de-açúcar e, em mistura com o glifosato, dessecou os perfilhos principais. Em pré-plantio incorporado, o óleo fúsel não apresentou interferência sobre a germinação e viabilidade das sementes; em pósemergência, os resultados mostraram controle parcial com a mistura do óleo fúsel e glifosato. Na pós-emergência tardia, em mistura com glifosato apresentou vantagens, por reduzir a massa seca das plantas e custos com a aplicação. / Abstract: The studies aimed to evaluate the fusel oil efficiency in sugarcane desiccation and weed control. The first study evaluated the efficacy of Energic, neutral detergent, alcohol and Haiten in fusel oil and water mixture. The second study evaluated the sugarcane desiccation compared with rates of fusel oil and glyphosate. The experimental design was completely randomized with 12 treatments and four replications in 22 L pots. At other study, isolated fusel oil was applied in incorporate preplanting of weeds, in experimental design completely randomized with 25 treatments and four replications analysed in a factorial arrangement 5x5 (concentrations and weed species); in weed post-emergence there were the fusel oil mixture with glyphosate, in different concentrations. The isolated fusel oil efficiency was evaluated and in mixture with glyphosate, applied in weed late post-emergence, in randomized blocks with 13 treatments in four 4 replications. The fusel oil mixture with water, Energic and neutral detergent provided better results, while alcohol and Haiten showed inefficient. The isolated fusel oil didn’t promote the complete sugarcane desiccation and when in mixture with glyphosate desiccated the sugarcane main shoot population. In incorporate pre-planting, the fusel oil didn't present interference about the seed germination and viability; in weed late post-emergence, the results showed partial control with fusel oil and glyphosate mixture. In weed late post-emergence, fusel oil mixture with glyphosate presented advantages, for reducing the plant dry mass and costs with the application. / Orientador: Marcos Omir Marques / Coorientador: Maria do Carmo Morelli Damasceno Pavani / Banca: Luciano Soares de Souza / Banca: Maria do Carmo de Salvo Soares Novo / Banca: Miguel Ângelo Mutton / Banca: Pedro Luís da Costa Aguiar Alves / Doutor

A parede celular vegetal é composta primariamente de celulose, hemicelulose e lignina. A lignina é o segundo biopolímero mais abundante na biosfera, atrás apenas de celulose, e é formado principalmente a partir da ligação entre três monômeros chamados monolignóis. A formação desses monolignóis é catalisada por pelo menos 10 enzimas membros das famílias gênicas AMP_binding (gene 4CL), p450 (C3H, C4H e F5H), ADH_N (CAD), Epimerase (CCR), Methyltransf_3 (CCoAOMT), Methyltransf_2 (COMT), Transferase (HCT) e Lyase_aromatic (PAL), que compõem a via de biossíntese dos monolignóis. Até o momento, cerca de 25 sequencias da via de biossíntese de lignina já foram identificados em cana-de-açúcar (Saccharum spp). Ainda, o sequenciamento do genoma desta espécie se mostra uma difícil tarefa devido ao tamanho do genoma e ploidia nuclear. Em virtude da disponibilidade de transcriptomas oriundos de diferentes órgãos de cana-de-açúcar e a importância na identificação correta das sequencias ortólogas, o presente trabalho visa à identificação, anotação e análise filogenética dos genes das famílias gênicas envolvidas na biossíntese de lignina em cana-de-açúcar. Para isso, genes destas famílias gênicas da planta modelo de eudicotiledôneas Arabidopsis thaliana e gramíneas tais como Oryza sativa (arroz), Brachypodium distachyon, Zea mays (milho) e Sorghum bicolor (sorgo) foram identificados, anotados e filogeneticamente categorizados. Em seguida, as sequências de cana-de-açúcar foram identificadas em cinco transcriptomas distintos, anotados e avaliados quanto à identidade e cobertura em relação às proteínas de sorgo. As análises filogenômicas entre cana-de-açúcar e as demais espécies estudadas revelaram 23 sequências candidatas envolvidos na biossíntese de lignina em cana-de-açúcar. / Plant cell wall is composed of cellulose, hemicellulose, and lignin. Lignin is the second most abundant biopolymer on biosphere, after only cellulose, and is formed primarily from three monomers, called monolignols. Monolignol biosyntesis is catalysed by at least 10 enzymes members of AMP_binding (4CL enzyme), p450 (C3H, C4H e F5H), ADH_N (CAD), Epimerase (CCR), Methyltransf_3 (CCoAOMT), Methyltransf_2 (COMT), Transferase (HCT) e Lyase_aromatic (PAL) gene families involved in the biosynthetic pathway of monolignols. So far, there is 25 sugarcane (Saccharum spp.) genes identified involved in the lignin biosynthesis. Sequencing sugarcane\'s genome is still a difficult task due to large genome and ploidy level. Due to the availability of transcriptome data from distinct organs of sugarcane and the importance in the accuracy on the identification of the orthologous sequences, this work aim the identification, annotation and phylogenetic analysis of proteins members of distinct gene family involved in monolignol biosynthesis in sugarcane. To conduct it, we identified and annotated all genes from the model plant eudicots Arabidopsis thaliana and grasses such as Oryza sativa (rice), Brachypodium distachyon, Zea mays (mayze) and Sorghum bicolor (sorghum) and analysed the phylogenic relationship among them. The sugarcane sequences were retrieved and annotated from five distinct transcriptome data and analysed by the identity and coverage against the best sorghum orthologous genes. The phylogenomics analysis revealed a total of 23 sugarcane sequences putatively involved in the biosynthesis of lignin.

O objetivo deste trabalho foi caracterizar a farinha do bagaço da cana-de-açúcar (FBCA) e avaliar o efeito de sua aplicação, como fonte de fibra insolúvel, em bolo. A FBCA foi caracterizada com relação a sua composição química (teor de umidade, cinzas, proteínas, carboidratos, fibras solúveis, fibras em detergente ácido e neutro e lipídeos), atividade de água (Aw), composição de minerais pela técnica de Espectrometria de Fluorescência de Raios-X (TXRF), determinação do tamanho e distribuição das partículas, avaliação da cor instrumental e contagem de bolores e leveduras e de coliformes totais. A FBCA foi aplicada em bolo tipo cupcake nas proporções de 3 e 6%. Os bolos recém-preparados foram avaliados com relação à umidade, atividade de água (Aw), densidade, cor instrumental e pH. Os bolos foram estocados durante 42 dias e em 5 pontos deste período foram avaliados com relação a umidade, Aw, perfil de textura (parâmetros dureza, elasticidade e coesão), contagem de bolores e leveduras e aceitação sensorial (atributos aparência, sabor, aroma, textura e aceitação global). Todas as análises foram realizadas também em uma amostra controle, preparada sem a adição de FBCA, para efeito de comparação. A FBCA demonstrou ser excelente fonte de fibra insolúvel, com 81,92% entre celulose, lignina e hemicelulose, bem como de minerais como o ferro, manganês e cromo. Todavia é uma fonte pobre de fibra solúvel, proteína, carboidratos e lipídeos. Em torno de 60% das partículas da FBCA possuem tamanho inferior a 80µm. A FBCA apresentou contagem alta de bolores e leveduras e cor tendendo para o amarelo e vermelho. Os bolos apresentaram em torno de 15% de umidade e Aw na faixa intermediária. Bolos com FBCA foram ligeiramente mais úmidos, escuros e densos que o controle. Porém, foram menos elásticos e tão coesos e duros quanto. Os bolos adicionados de 3% de FBCA apresentaram a mesma aceitação que o produto tradicional, com a vantagem do maior valor nutricional em função da presença das fibras e dos minerais oriundos da FBCA. / The aim of this study was to characterize the flour from sugarcane bagasse (FSB) and to evaluate the effect of its application as a source of insoluble fiber in cake. The FSB was characterized in respect to the chemical composition (moisture content, ash, proteins, carbohydrates, soluble fiber, fiber in acid and neutral detergent, cellulose, hemicellulose, lignin and lipids), water activity (Aw), mineral composition by Fluorescence spectrometry technique X-ray (TXRF), size and distribution of particles, evaluation of the instrumental color and counts of molds and total coliforms. The FSB was applied in cupcakes, in proportions of 3% and 6%. The freshly prepared cakes were evaluated in respect to moisture content, water activity (Aw), density, pH and instrumental color. The cakes were stored for 42 days and during this period were evaluated for moisture content, Aw, instrumental texture (hardness, elasticity and cohesion), counts of molds and sensory acceptance (appearance, taste, flavor, texture and overall acceptance). All analysis were also carried out in a control sample, prepared without the addition of FSB, for comparison. The FSB demonstrated to be an excellent source of insoluble fiber, containing 81.92% between cellulose, lignin and hemicellulose, and is source of minerals such as iron, manganese and chromium. However, it is a poor source of soluble fiber, protein, carbohydrates and lipids. Around 60% of the particles of FSB are smaller than 80µm. The FSB presented high counts of molds and the color tends to yellow and red. The cakes showed around 15% of moisture content and Aw in the middle range. Cakes with FSB were slightly damper, darker and denser than traditional. However were less elastic, but so cohesive and hard as the traditional one. Cakes added with 3% of FSB presented the same acceptance that the traditional product, with the advantage of greater nutritional value due to the presence of fibers and minerals from FSB.

A fermentação de hidrolisados hemicelulósicos tem como principal dificuldade a presença de compostos inibidores ao metabolismo microbiano, derivados da degradação parcial da lignina, degradação dos açúcares e liberação de radicais acetil durante a etapa de hidrólise dos materiais lignocelulósicos. O presente trabalho teve por objetivo avaliar a influência de diferentes tratamentos de destoxificação sobre a composição e fermentabilidade do hidrolisado hemicelulósico de bagaço de cana-de-açúcar na produção de etanol e xilitol. Foram avaliados os seguintes tratamentos: a) alteração de pH com óxido de cálcio e ácido fosfórico seguido de adsorção com carvão ativo; b) utilização de resinas de troca iônica (A-860, A-500PS e C-150 - Amberlite); e c) extração líquido-líquido com interface imobilizada em membrana de fibra oca (Membrana, Charlote, NC - USA), na qual a fase orgânica foi uma mistura de octanol e Alamina 336 e a fase aquosa foi o próprio hidrolisado. De acordo com os resultados, os tratamentos aplicados (a, b e c) promoveram uma redução na concentração de ácido acético de 46,74%, 64,15% e 44,71% e uma redução na absorbância relativa (A.R.) de 82,0%, 94,59% e 46,07%, respectivamente. Na fermentação dos hidrolisados tratados, pela levedura Candida guilliermondii FTI 20037, após 48h de fermentação, os resultados para o fator de rendimento foram de 0,57g/g, 0,42g/g e 0,33 g/g após os tratamentos a, b e c, respectivamente. Quanto à produtividade volumétrica, na mesma ordem dos tratamentos foram obtidos os seguintes resultados: 0,39 g.L-1.h-1, 0,20 g.L-1.h-1 e 0,16 g.L-1.h-1. Nas fermentações dos hidrolisados tratados (a) e (b) pela levedura Pichia stipitis IMH 43.2, visando a produção de etanol, após 48h os resultados do fator de rendimento foram 0,38 g/g e 0,23 g/g, respectivamente, enquanto a produtividade volumétrica foi de 0,09 g.L-1.h-1 para o tratamento (a) e para o tratamento (b). Para o tratamento (c) não se observou a produção de etanol nas condições de fermentação utilizadas. Considerando os resultados obtidos, sugere-se que, para as condições experimentais empregadas, o hidrolisado seja tratado por alteração de pH seguido de tratamento com carvão ativado, uma vez que este tratamento resultou nos melhores resultados para o fator de rendimento tanto na produção de xilitol quanto na produção de etanol. / The fermentation of hemicellulosic hydrolysates has as main difficult the presence of compounds derived from partial degradation of lignin, sugars and release of acetyl groups, which are inhibitors of microbial metabolism, during the hydrolysis process of lignocellulosic materials. This study aimed to evaluate the influence of different detoxification treatments on the composition and fermentability of the sugarcane bagasse hemicellulosic hydrolyzate in the production of xylitol and ethanol. The following treatments were evaluated: a) change of pH with calcium oxide and phosphoric acid followed by adsorption with activated charcoal, b) use of ion exchange resins (A-860, A-500 PS and C-150 - Amberlite) and c) liquid-liquid extraction with immobilized interface in hollow-fiber membrane (Membrana, Charlote, NC - USA), in which the organic phase was a mixture of octanol and Alamine 336, and the aqueous phase was the hydrolyzate. According to the results, the employed treatments (a, b and c) promoted a reduction of 46.74%, 64.15% and 44.71% in the acetic acid concentration, and a reduction in the relative absorbance (RA) of 82.0%, 94,59% and 46.07% respectively. In the fermentation of the treated hydrolyzates by the Candida guilliermondii FTI 20037 yeast, aiming xylitol production, after 48h of fermentation, the results for xylitol yield were 0.57g/g, 0.42g/g and 0.33g/g, after the treatments a), b) and c), respectively. The volumetric productivity obtained was 0,39 g.L-1.h-1, 0,20 g.L-1.h-1 and 0,16 g.L-1.h-1, in the same order of treatments. In fermentation by Pichia stipitis IMH 43.2 of the hydrolyzates that used treatments (a) and (b), aiming the production of ethanol, after 48h, the results for yield were 0.38g/g and 0.23g/g, respectively, while the volumetric productivity was 0,09 g.L-1.h-1 for both treatments. For treatment (c) there was no ethanol production using the fermentation conditions applied in this work. Considering these results, it is suggested that for the experimental conditions used at this work, the hydrolyzate should be treated by pH change followed by treatment with activated charcoal, since this treatment resulted in better results of yield in the production of xylitol and ethanol.