Programa de Pós-graduação em Genética e Melhoramento de Plantas
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Item Pré-melhoramento genético, floração in vitro e criopreservação de orquídeas nativas do cerrado(Universidade Federal de Goiás, 2014-11-27) Carneiro, Luciano Lajovic; Faria, Ricardo Tadeu de; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Carvalho, Virgínia Silva; Galdiano Junior, Renato Fernandes; Ferreira, Wagner de Melo; Santos, Izulmé Rita ImaculadaThe vast majority of orchids species are pollinated by insects, mainly bees, butterflies and moths. The orchid family is generally considered allogamic, but information about the type of reproduction are unknown for most species. Little information exist about orchids of Cerrado, which hinders plant breeding of these species and postpones their use in developing new varieties for the ornamental market. Recent methods of plant biotechnology can assist both plant breeding and conservation of these species, such as induction of early in vitro flowering as important technique particularly for species with a long cycle, and cryopreservation as a tool for the conservation of genetic variability. This study aims to contribute to the improvement and conservation of orchids with distribution in the Brazilian Cerrado. Selfcompatibility was evaluated for seven species: Cohniella cepula, Cyrtopodium eugenii, C. saintlegerianum, Epidendrum densiflorum, Epidendrum nocturnum, Epidendrum secundum e Lockhartia goyazensis. Only three were autocompatible: Epidendrum nocturnum, Epidendrum secundum and Lockhartia goyazensis. Interspecific compatibility was assessed for Cyrtopodium and Epidendrum genera, using reciprocal crosses. Just E. nocturnum and C. eugenii produced viable seeds. For early in vitro floral induction 20 different treatments were used through nutritional changes and use of cytokinin (BAP) in modified MS medium. Only C. cepula showed positive responses to treatment with formation of 46% floral stems. The effect of BAP was evident in the results. Cryopreservation was tested by vitrification of seeds of seven species of orchids, obtained from outcrossing. The responses to cryopreservation were partially different for each species. Most seeds tested showed any significant differences between freezing in liquid nitrogen without cryoprotectants and control without freezing. The freezing without cryoprotectant was successful to maintaining seed viability for all species, C. cepula 63%, C. eugenii 59%, C. sainttlegerianum 70%, E. densiflorum 42%, E. nocturnum 31%, E. secundum 69% and L. goyazensis 52%. Only E .nocturnum and C. cepula showed a significant reduction in viability when submitted to freezing in liquid nitrogen. The results presented here are useful for the development of breeding programs for orchids and conservation of these species.Item Isolamento de bactérias endofíticas e estabelecimento in vitro de diferentes genótipos(Universidade Federal de Goiás, 2012-08-10) Faria, Paulo Roberto; Araújo, Leila Garcês de; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Carneiro, Maurízia de Fátima; Vieira, José Daniel GonçalvesIn micropropagation, plant tissue fragments called explants are isolated aseptically, disinfected and cultured in culture medium. However, oxidation and contamination are the main problems in the initiation of the tissue culture plants. In order to initiate the establishment of in vitro methodologies for selected sugarcane varieties a series of decontaminating treatments were performed in shoot tips with different exposure times and concentrations of sodium hypochlorite, casugamicin, ethanol and mercuric chloride. There were no problems with oxidation in any treatment. The use of reagent concentrations and times tested showed not be phytotoxic. Treatment using longer times of exposure to casugamicin and sodium hypochlorite obtained the lowest levels of contamination by fungi and bacteria. However, bacterial contamination rates were high which indicates the necessity of using new procedures for the decontamination process. It is known that the sugarcane is associated with different endophytic bacteria. In order to better study these endophytic bacteria and their relationship with the host plant, promoted the isolation of bacteria in explants of five different varieties of sugarcane: RB98710, RB34068, RB034130, RB034120 and RB034041 to determinate of morphological and physiological characteristics. The thirteen isolates have diversity for morphological and biochemical staining and are characterized as gram-positive. All of them had a production capacity of plant growth factors, and after antimicrobial susceptibility testing, two antibiotics: Ciprofloxacin and Ceftazidime showed the greatest potential for controlling the growth of the isolates. With microbial contamination controlled, studies were conducted to evaluate the effect of growth regulators on in vitro multiplication of shoot tip culture and its roots in three sugarcane varieties: RB98710, RB034041 and RB034130. We used liquid MS medium containing BAP, KIN, NAA and GA3 in different combinations and concentrations. The results showed that, for the three varieties tested, MS medium with 0.2 mg.L-1 BAP and 0.1 mg L-1 KIN are most suitable for higher production of shoots and that the best rooting results were obtained in semi-solid ½ MS medium with 6% sucrose, 5.0 mg L-1 NAA and 0.2% activated charcoal. The plantlets were acclimatized and survivability under ex vitro conditions was 90%, three weeks after the transfer to the greenhouse.Item Estabelecimento in vitro de Handroanthus impetiginosus (Mart. ex Dc.) Mattos (Bignoniaceae) e estudo da incidência de oídio (Oidium sp.) em plântulas obtidas in vitro(Universidade Federal de Goiás, 2013-04-02) Mamedes, Talita Cristina; Araújo, Leila Garcês de; http://lattes.cnpq.br/3488880262260757; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Fillippi, Marta cristina Corsi de; Gonçalves, Letícia de AlmeidaHandroanthus impetiginosus (Mart. ex DC.) Mattos is widely distributed woody species in Brazil. It has economic importance for its use as wood, medicinal, ecological and ornamental tree. There is great variation in the production of fruits and seeds over the years, many of the seeds are attacked by fungi and insects, and finally, the remaining intact seeds lose viability very quickly. Techniques of plant tissue culture can increase the rate of germination in woody species. The cultivation in nutrient medium under aseptic conditions, provides great production of seedlings in reduced time and space and at any time of year. Furthermore, in vitro propagation acts as a tool in the identification of microorganisms, studying their relationships and interactions with the host plant at genetic, cellular and physiological levels. The aim of this study was to establish a protocol for in vitro propagation of H. impetiginosus developed in three steps: decontamination and seed germination, induction of new shoots and roots. The protocol was based on direct organogenesis using plant material from seedlings germinated in vitro. For in vitro germination, seed surface contamination was controlled using 2 min. in 70% ethanol and 10 min. sodium hypochlorite with 2 % active chlorine. The germination rate was 99 % on MS medium. Induction and multiplication of new shoots were carried out with the aid of growth regulators 6 - benzylaminopurine (BAP) and naphthalene acetic acid (NAA) in different combinations. The results showed that for explants of H. impetiginosus all the tested amounts of BAP and NAA have the ability to regenerate shoots, callus and adventitious roots, it is necessary to improve the regeneration protocol to obtain greater number of new shoots. For in vitro rooting, cuttings responded positively to 6.0 mg L - 1 butyric acid (IBA), with 92 % rooting. However, the methods of decontamination of seeds did not control fungi of the genus Oidium. In order to identify the fungus and study the fungus - host relationship in H. impetiginosus cuts were made in vitro leaf fragments and the anatomical structure analyzed by optical microscope and scanning electron microscope. After the emergence of Oidium sp. on seedlings developed in vitro, methods of seeds disease control were tested for germination using different times and concentrations of ethanol, sodium hypochlorite, Chlorothalonil + tiophanate Methyl (systemic fungicid ) and Neem oil assessing the incidence and severity of each disease treatment. No treatment was effective in the control of powdery mildew, indicating the need for further studies. However, reduction of severity was observed in seeds treated with 70 % ethanol and sodium hypochlorite (2% active chlorine) and Neem oil immersion to 1.5 % for 10 minutes.Item Cultivo in vitro e citogenética de Cyrtopodium saintlegerianum Rchb. f.(Orchidaceae: Cyrtopodiinae)(Universidade Federal de Goiás, 2012-10-30) Silva, Daniella Mota; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Takane, Roberto Jun; Faria, Ricardo Tadeu deCyrtopodiumsaintlegerianumRchb. f is an epiphytic species typical of the Midwest, especially in distributed Brazilian Central Plateau, and has a wide geographical distribution in Brazil. It is usually found in the trunks of palm trees, forming large clumps. It has good features for ornamentation, for the beauty and size of its inflorescence, however, are not found in the literature about its conservation, methods for its spread or that could be used in floriculture and landscaping. Thus, this study aimed to establishing protocols for germination asymbiotic effects of phytohormones and acclimatization and characterization of chromosomal species. In 2010, the establishment of micropropagation protocols, capsules were collected in a pasture area in the municipality of Mossâmedes, GO, then the part previously sterilized seeds were separated and tested in a 1% tetrazolium dye.. For cultivation was tested asymbiotic different culture media and then tested after different concentrations and combinations in treatments BAP 16 / ANA, to finally evaluate the acclimatization substrates combined and fertilized with chemical fertilizers and organic. For the karyotype of the species, the plant material is derived from plants grown in vitro in culture medium. We tested four protocols, with differences in enzyme solution for softening the roots, dyes, anti-mitotic concentration of the solution and hydrolysis, and the use of growth regulators for root induction in vitro. All protocols were in common roots pretreated with anti-mitotic 8-hydroxyquinoline (0.002 M) in refrigerator for 24 hours. Then protocols roots were fixed in Carnoy 3:1 for 18 hours the first and second and third and fourth protocol for 24 hours at room temperature. After stored at -20 º C in the same fixative for further analysis (only the fourth protocol). The roots of the protocols were stained with different dyes: hematoxylin, Schiff, acetic orcein and Giemsarespectively.The results of germination was satisfactory in all culture media. For medium supplemented with auxin / cytokinin combined the best concentrations for variable height were 0.2 mg L-1 NAA and BAP without adding control without addition of regulators. The best means to induce large numbers of shoots were 4 mg L-1 BAP and 4 mg L-1 BAP / 0.2 mg L-1 NAA. The number of leaves was rated best in the concentrations of BAP without NAA at concentrations of 1.0, 2.0 and 4.0 mg L-1. The treatments with the highest number of roots were control without added growth regulators at doses of 0.2, 0.5, 1.0 mg L-1 NAA without addition of BAP, as well as the length of roots was favored by the same treatments. The largest number occurred in callus treatment with concentrations of 1.0 mg L-1 BAP without adding ANA. ForcytogeneticsC. saintlegerianum the best protocol was evaluated with the regulator which was obtained metaphases, however chromosomes were condensed, and the number of chromosomes was found to be 2n = 48. cides.Item Germinação, estabelecimento e multiplicação in vitro de Eugenia dysenterica DC. e Dipteryx alata Vogel, espécies frutíferas do cerrado(Universidade Federal de Goiás, 2012-08-09) Silva, Lívia Cristina da; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Ferreira, Luciano Domingues Bittencourt; Gonçalves, Letícia de AlmeidaDipteryx alata Vogel and Eugenia dysenterica DC. are Cerrado’s fruit tree threatened by habitat fragmentation and the predatory extractivism. Thus, it is essential to the study of techniques for the conservation and sustainable use of these species. The objective for this work was to establish protocols for micropropagation of these species from the in vitro germination of their seeds. Experiments were conducted at the Laboratory of Plant Tissue Culture ICB / UFG. Seeds of both species were divided into two groups: with coat and without coat. After pre-cleansing with detergent and alcohol 70%, seeds of D. alata were treated with four concentrations of sodium hypochlorite. Seeds of E. dysenterica were treated with four concentrations of sodium hypochlorite and three of casugamicina. The seeds were inoculated in ½ MS and MS complete, with or without addition of charcoal. The lowest contamination percentage for E. dysenterica occurred with seeds without tegument soaked in 0.5% of active chlorine. To D. alata, the most effective treatment was with seed-coats soaked in 1.25% of active chlorine. For the germination of E. dysenterica, the seed coats were removed. The treatments used through complete MS and ½ MS. After inoculation, the seeds remained in a growth chamber in two distinct photoperiods: 16 h light or 24 hours of dark. The highest germination percentage for E. dysenterica, with 93%, occurred in complete MS medium in a 16h photoperiod. To D. alata, the highest germination percentage, 97.5%, occurred in complete MS medium without charcoal, in a 16h photoperiod. To verify the induction of shoots of both species, nodal segments were inoculated on MS medium supplemented with different concentrations of NAA and BAP, the best treatment for multiplication of shoots in E dysenterica was with 4.0 mg.L-1 BAP. For D. alata, the best was 0.1 mg.L-1 NAA and 2.5 mg l-1 BAP. To study the roots and shoots from in vitro cultures inoculated in a MS or ½ MS supplemented with combinations of NAA, sucrose and activated charcoal. No satisfactory results occurred for rooting in any species. For E. dysenterica, the few seedlings that emitted roots did not stand the acclimatization. To D. alata, no emission of roots was detected, but there was the issue of shoots in all treatments, especially those inoculated in ½ MS. To obtain the callus, stem explants of D. alata, and leaf explants of E. dysenterica were inoculated on MS medium supplemented with different concentrations of BAP and 2.4 D. As a result, we obtained callus formation in D. alata with BAP, ranging from 0.0 mg L-1 to 1.0 mg.L-1, interacting with 2,4-D, ranging from 1.0 mg L-1 and 4.0 mg L-1. Leaf explants of E. dysenterica callus was obtained between 3.0 and 4.0 mg.L-1 of 2.4-D combined with 0.5 and 1.0 mg.L-1 BAP.Item Micropropagação de Bambusa oldhamii Munro e biocaracterização de fungos endofíticos multifuncionais(Universidade Federal de Goiás, 2018-07-20) Silveira, Andreia Alves da Costa; Araújo, Leila Garcês de; http://lattes.cnpq.br/3488880262260757; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Lanna, Anna Cristina; Carrer Filho, Renato; Fillipi, Marta Cristina Corsi de; Gonçalves, Fábio JoséIn micropropagation, seedlings are produced in large scale with homogeneity, which can be potentiated with photomixotrophic systems, which improve the physiological quality of the seedlings. Endophytic fungi of bamboo have the potential of producing substances that can promote biofertilization and antagonism against pathogens. An example of application is antagonism against Magnaporhe oryzae, considered the main pathogen of rice. The objective of this work was to establish the micropropagation protocol of Bambusa oldhamii; besides characterizing endophytic fungi of this species, verify the biofertilizer potential in rice and potential antagonist against M. oryzae. Buds were inoculated in MS medium supplemented with 0.45 μM Tidiazuron-TDZ, 0.2% (w / v) PPM (Plant Preservative Mixture) and 50 mg L-1 kanamycin. The treatments consisted of: different months of collection (March-December 2016) x types of cap (heterotrophic and photomixotrophic system) x luminous conditions (100% blue (455nm), 100% red (630nm), 30% blue + 70% For the multiplication, shoots were inoculated in MS medium + 2.27 μM TDZ or 3.40 μM Paclobutrazol - PBZ. The fungal isolates that were observed in the in vitro culture were identified and characterized biochemically, besides antagonism to M. oryzae and microbiolization of rice seeds, June and July were constituted in the best months of collection, and the climatic variables that most interfered in the morphogenesis were minimum temperature and compensated mean. Photomixotrophic system was superior in the multiplication with TDZ, with increase of 59.51% in 30% blue + 70% red, 70.80% in 30% red + 7 0% blue, and 50.49% in white fluorescent lights. PBZ was higher in carotenoid production, with a mean of 128.02 μg / mL in blue light and conventional lids. TDZ was higher than PBZ when blue was not present. Five potential fungal isolates were identified, 29 (Acrocalymma sp.), 122 (Botryobambusa fusicoccum) 711 (Phoma sp.), 712 (Phoma sp.) And 27 (Arthrinium marii) isolates 711, 712, 27 and 29 produced PPO. The isolate 29 produced higher amounts of AIA, with 31.55 mg / mL on the fourth day. It was observed a reduction of mycelial growth of M. oryzae by all isolates, with emphasis on isolate 122. Isolate 711 presented phosphate solubilization, and higher mean shoot length, fresh and dry mass in rice microbiolization. An efficient micropropagation protocol of B. oldhamii using a photomixotrophic system with 30% blue + 70% red LEDs could be established, as well as the identification of a fungal isolate promising for biofertilization, which could be the basis for new studies with rooting of B. oldhamii and promotion of growth in newly acclimatized species.Item Criopreservação de ápices caulinares e micropropagação em condições heterotróficas e mixotróficas de eugenia dysenterica (Mart.) DC.(Universidade Federal de Goiás, 2015-02-23) Silveira, Andreia Alves da Costa; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Oliveira, Saulo Araújo de; Gonçalves, Letícia de AlmeidaConventional micropropagation systems often use culture media supplemented with sucrose and sealed bottles. This system is called heterotófico as exogenous carbohydrates are the only source of energy for the plant. Mixotróficos systems, although still adding sucrose in half, have the differential to allow gas exchange between the internal and external environment of the culture flask. This ventilation brings many benefits to plant growth in vitro. By the constant loss of the Savannah area, it is necessary to improve micropropagation protocols and germplasm conservation of the species within it. Cryopreservation is considered the best for long-term preservation method and the encapsulation-dehydration technique have been promising for tropical species. The objectives were to compare the heterotrophic system in vitro propagation of Eugenia dysenterica (Mart.) DC. mixotrófico with a system, and get a method of germplasm storage dysenterica E. vitro by cryopreservation. Identified Gems' positions in the stem (closer to the root or the apex) were inoculated in tubes with or without gas exchange in WPM medium supplemented with 0,00μΜ, 0,54μM, 2,69μM, and 5,37μM, NAA, and 0,00μM, 4,44μM, 11,10μM and 17,76μM BAP in all possible combinations. For rooting was used WPM medium supplemented with IBA to 0,00μM, 9,84μM, 19,68μM, and 29,52μM. The system of gas exchange was greater in the number of sheets in most analyzed variables. The type of gem had little influence on the variables. The best treatment in heterotrophic and mixotrófico systems was 29.52 mM of IBA, with 33.33% and 16.66% respectively rooting. Acclimatization was performed successfully in both systems. For cryopreservation was used apexes obtained from plants grown in vitro. The apices were initially suspended in MS medium supplemented with 3% solution of sodium alginate, 0.4 M glycerol and 4,44μM BAP and dispensed in MS medium solution (free of calcium) supplemented with 0.1 M calcium chloride, 0.4 M glycerol and 4,44μM BAP. The summits were held in three exposure levels of sucrose (0.25M, 0.5M, and 0.75M) combined will three levels of desiccation (0 h, 1 h, 2 h) before being frozen in liquid nitrogen. There was no regeneration in cryopreserved apexes. The best treatment for non-cryopreserved apices consisted of 0.25M sucrose and 1 h of drying. The null regeneration of cryopreserved apices which indicates that the stress is related to desiccation stress freezing.Item Propagação vegetativa por alporquia e otimização de protocolo de estabelecimento in vitro de mangabeira (Hancornia speciosa Gomes)(Universidade Federal de Goiás, 2020-02-28) Tiago, Bruno dos Santos; Sibov, Sérgio Tadeu; http://lattes.cnpq.br/4627553641870284; Sibov, Sérgio Tadeu; Ganga, Rita Maria Devós; Aguiar, Renata Alves deHancornia speciosa Gomes, popularly known as mangabeira, is a native fruit species of the Cerrado used for food purposes and as herbal medicine. Currently, due to habitat loss and predatory extraction, there is a great demand in more studies for the conservation and use of this fruit species. The main challenge for the in vitro establishment of the species is the high number of endophytic agents (fungi and bacteria) that are present in plant tissues. This makes it more difficult to introduce plant material into the in vitro culture. This work aimed to optimize the in vitro establishment protocols in addition to carrying out vegetative propagation, via layering, generating clones of matrix plants. For the in vitro establishment of one of the varieties of the species, var. gardineri, first the fall fruits were collected in the Collection of Native Fruits of the Cerrado of the School of Agronomy at UFG. Fruits were pulped and 80 seeds were selected, which had their tegument removed and proceeded to the decontamination process. In a laminar flow chamber, two groups of seeds were separated: 40 seeds were opened and their embryos exposed, isolated and inoculated in MS culture medium; the other 40 seeds were inoculated directly in MS culture medium. To test the effect of stretching on reducing the period of development in vitro, embryos were inoculated in treatments with 0, 1, 2 and 4 μM of gibberellic acid (GA3) and later in a second experiment in MS medium with 1,0 μM of 6-benzylaminopurine (BAP), with lower concentrations of GA3: 0.2; 0.4; 0.6; 0.8 and 1 μM. For the next stage, treatments were carried out to root the new shoots with indolbutyric acid (IBA) in concentrations 0, 1, 3, 5 and 7 mg.mL-1 . The germination of the isolated embryos showed less than 5% of contamination and was 30% faster when compared to the seed. The effect of GA3 on isolated embryos allowed the plant to elongate with 0.8 μM, reaching 130 mm in 75 days, reducing in vitro cultivation by more than 100 days. In rooting, the concentration of 1 mg.mL-1 of AIB allowed more than 9 roots per plant and an average length of 54 mm. The acclimatization of the treatment plants took place on substrate with 50% sand and 50% broad red soil with commercial substrate. In this process, mortality was less than 10%. The layering experiments were carried out with four varieties of H. speciosa: gardineri, speciosa, cuyabensis and pubescens and in two different seasons: the dry season, and the rainy season. AIB was used to induce rooting at concentrations of 0, 4, 5 and 6 mg.mL-1 , with 15 ml per branch. The substrate used was soil humidified with commercial substrate fixed to the ringed branch. After 180 days, the best results were in the concentration of 5 mg.mL-1 in the four varieties used. However, the results for each variety were at different times of the year, due to the specific phenological phases.