ABSTRACT Although karyotype features are useful data for evolutionary studies, cytogenetic data in Bambusoideae are mainly based only on chromosome counts. The first comparative cytogenetic analysis of three Neotropical woody bamboo species - Guadua chacoensis, G. angustifolia and Chusquea tenella - was undertaken based on new and reviewed chromosome counts, CMA/DAPI double staining, fluorescent in situ hybridization (FISH) with 35S and 5S rDNA probes, and genome size estimation. The two species of Guadua were found to have 2n = 46 chromosomes, while the first record for C. tenella was 2n = 44. Only one pair of CMA+/DAPI- was detected on the terminal region of metacentric chromosomes in all three species. Likewise, one pair of 5S and 35S rDNA sites was detected in all three species, with the 35S rDNA sites always collocated with the CMA+ bands. Genome sizes ranged from 2C ≈ 3.99 pg for the species of Guadua, to 2C = 4.77 pg for C. tenella. Considering the Miocene origin of Neotropical woody bamboos, the observed karyotype stability suggests that the analyzed species are diploidized paleopolyploids. The results reveal the conservative cytomolecular organization of Neotropical woody bamboo karyotypes, which helps to improve our understanding of the evolution of this group.

ABSTRACT Eugenia uniflora is an American tree species with ecological, agronomical and medicinal importance. However, just few studies have focused on the in vitro propagation and production of secondary metabolites. This study investigated the explant sources and culture conditions for the in vitro callogenesis in E. uniflora towards induction of somatic embryogenesis and production of antioxidant compounds. Nodal segments, leaf sections and root segments from in vitro germinated seeds were used as explants and eight different combinations of auxins (2,4-D or NAA) and cytokinins (BAP or TDZ) were tested for the callus induction. The best callogenic response was observed in nodal segments, followed by leaf sections. Root segments presented comparatively poorer callogenic performance. Calli from nodal segments cultivated in MS medium with NAA (10 µM) + TDZ (5 µM) originated putative pro-embryogenic structures, while the culture in liquid medium using MS supplemented with NAA (10 µM) + TDZ (5 µM) revealed significantly higher content of phenols and flavonoids, as well as higher reducing capacity than the tested treatments and the control (fresh leaves). In summary, the calli obtained from nodal segments revealed competence for somatic embryogenesis induction and development as well as the production of secondary metabolites with pharmaceutical potential.

RESUMEN Eugenia uniflora es una especie arbórea americana de importancia ecológica, agronómica y medicinal. Sin embargo, pocos estudios se han centrado en la propagación y producción in vitro de metabolitos secundarios. Este estudio investigó las fuentes de explantes y las condiciones de cultivo para la callogénesis in vitro en E. uniflora para la inducción de embriogénesis somática y la producción de compuestos antioxidantes. Se utilizaron segmentos nodales, secciones de hojas y segmentos de raíz de semillas germinadas in vitro como explantes y se probaron ocho combinaciones diferentes de auxinas (2,4-D o ANA) y citoquininas (BAP o TDZ) para la formación de callos. La mejor respuesta callogénica se observó en segmentos nodales, seguidos de secciones foliares. Los segmentos de raíz presentaron un rendimiento callogénico comparativamente menor. Los callos de segmentos nodales cultivados en medio de cultivo MS con NAA (10 µM) + TDZ (5 µM) originaron estructuras proembriogénicas putativas, mientras que el cultivo en MS medio líquido con ANA (10 µM) + TDZ (5 µM) reveló un contenido significativamente mayor de fenoles y flavonoides, así como una mayor capacidad reductora que los tratamientos probados y el control (hojas frescas). En resumen, los callos obtenidos de segmentos nodales revelaron competencia para la inducción y el desarrollo de embriogénesis somática, así como para la producción de metabolitos secundarios con potencial farmacéutico.

ABSTRACT. Tissue culture techniques have been employed for bromeliad mass propagation by means of the morphogenetic route of nodular cluster cultures (NCs). This study aimed to assess protein, carbohydrate and global DNA methylation (GDM) level dynamics during NCs induction from Vriesea reitzii seeds. Seeds were inoculated into Murashige and Skoog (MS) liquid medium supplemented with 4 µM α-naphthaleneacetic acid (NAA) to induce NCs and in culture medium without plant growth regulators to form normal seedlings. Samples collected at 0, 3, 7, 10, 14, and 21 days of culture were analyzed. All parameters assessed showed the same variation pattern. However, seeds inducing NCs showed significantly lower starch (6.0 mg g-1 FM), carbohydrate (10.7 mg g-1 FM) and GDM (11.0%) levels than seeds forming normal seedlings after 21 days in culture. On the other hand, the protein content (9.1 mg g-1 FM) was significantly higher during induction. NCs induction process through seeds is the result of gene reprogramming in the explant, which leads to morphological, biochemical and metabolic alterations. This involves dedifferentiation, high cell proliferation, high energy demand and protein synthesis, which is related to elevated metabolic activity.

Abstract Plastomes are very informative structures for comparative phylogenetic and evolutionary analyses. We sequenced and analyzed the complete plastome of Campomanesia xanthocarpa and compared its gene order, structure, and evolutionary characteristics within Myrtaceae. Analyzing 48 species of Myrtaceae, we identified six genes representing ‘hotspots’ of variability within the plastomes (ycf2, atpA, rpoC2, pcbE, ndhH and rps16), and performed phylogenetic analyses based on: (i) the ycf2 gene, (ii) all the six genes identified as ‘hotspots’ of variability, and (iii) the genes identified as ‘hotspots’ of variability, except the ycf2 gene. The structure, gene order, and gene content of the C. xanthocarpa plastome are similar to other Myrtaceae species. Phylogenetic analyses revealed the ycf2 gene as a promissing region for barcoding within this family, having also a robust phylogenetic signal. The synonymous and nonsynonymous substitution rates and the Ka/Ks ratio revealed low values for the ycf2 gene among C. xanthocarpa and the other 47 analyzed species of Myrtaceae, with moderate purifying selection acting on this gene. The average nucleotide identity (ANI) analysis of the whole plastomes produced phylogenetic trees supporting the monophyly of three Myrtaceae tribes. The findings of this study provide support for planning conservation, breeding, and biotechnological programs for this species.

Abstract Butia eriospatha is an endemic palm species from the Atlantic Rainforest in Brazil, a biodiversity hotspot. This species is currently listed in the IUCN red list as vulnerable and lacks specific plastid markers for population genetics studies. In addition, the evolutionary relationship within the genus Butia is not yet well resolved. Here, we sequenced and characterized the complete plastid genome (plastome) sequence of B. eriospatha. The complete plastome sequence is 154,048 bp in length, with the typical quadripartite structure. This plastome length and genes content is consistent with other six species from tribe Cocoseae. However, the Inverted Repeat (IR) borders show some variation among the analyzed species from this tribe. Species from the Bactridinae (Astrocaryum and Acrocomia) and Elaeidinae (Elaeis) subtribes present the rps19 gene completely duplicated in the IR region. In contrast, all plastomes sequenced from the subtribe Attaleinae (Butia, Cocos, Syagrus) present one complete CDS of rps19 and one partial copy of rps19. The difference in the IR/LSC junctions between Attaleinae and the sister clades Bactridinae + Elaeidinae might be considered an evolutionary signal and the plastome sequence of B. eriopatha may be used in future studies of population genetics and phylogeny.

Abstract MON810 is a genetically-modified (GM) maize (Zea mays) event commonly employed in insect-resistant GM maize hybrids. GM events obtained by biolistic transformation methods, such as MON810, are generated by insertion of a recombinant gene expression cassette in a random locus of the plant genome, and this process may cause emergent properties besides the intended modification. Here, we compared morphophysiological parameters of MON810 GM maize hybrid AG-5011YG and its non-GM near-isogenic hybrid (NIH) AG-5011, using in vitro cultures as an interactive model. NIH callogenesis frequency, callus friability, and de novo morphogenesis were compared using two explant types and different 2,4-dichlorophenoxyacetic acid (2,4-D) levels. 2,4-D modulated the morphophysiological responses of both NIHs, but when using root segments as explants, the GM showed significantly different trends for callus induction and friability, with lower responses at higher 2,4-D concentrations, indicating an emergent property related to altered cell response to 2,4-D.

RESUMO A cadeia produtiva de bambu, no Brasil, ainda é incipiente, e tem baixa oferta de mudas devido aos métodos convencionais de propagação. Objetivou-se, aqui, estabelecer um protocolo de micropropagação para Guadua chacoensis. Explantes de matrizes cultivadas em ambiente controlado resultaram em menor contaminação, relativamente aos de plantas cultivadas em campo. A contaminação foi ainda mais reduzida quando se adicionaram 2 mL L-1 de Plant Preservative Mixture (PPM™) ao meio de cultura, resultando em maior taxa de estabelecimento. As culturas foram multiplicadas por meio de segmentos nodais de plantas in vitro ou por divisão de touceiras em concentrações crescentes (0 µM, 10 µM, 20 µM, 30 µM ou 40 µM) de 6-Benzilaminopurina (BAP). O número de brotações aumentou com o incremento na concentração de BAP; porém, isto reduziu a taxa de enraizamento e o comprimento de raízes. Plantas aclimatizadas sob 0 %, 35 % ou 65 % de sombreamento apresentaram rendimento quântico máximo do fotossistema II (Fv/Fm) dinâmico, o qual diminuiu nos primeiros sete dias após a transferência para condições ex vitro e, em seguida, aumentou até atingir valores estáveis de 0,775, aos 17 dias. Além disso, o sombreamento aumentou as taxas de sobrevivência das plantas, em comparação ao tratamento sem sombreamento, o qual apresentou sintomas de fotoinibição e fotodano.

ABSTRACT The bamboo productive chain is still incipient in Brazil, and the low supply of plantlets due to low-efficient conventional propagation methods presents a significant bottleneck to its development. This study aimed to establish a micropropagation protocol for Guadua chacoensis. Explants from donor plants cultivated under controlled environment showed less contamination, if compared to explants from plants grown in the field. The contamination rate was even lower when 2 mL L-1 of Plant Preservative Mixture (PPM™) were added to the culture medium, leading to a higher establishment rate. The obtained cultures were then multiplied using either in vitro-derived nodal segments or clump division in the presence of increasing contents (0 mM, 10 mM, 20 mM, 30 mM or 40 mM) of 6-Benzylaminopurine (BAP). The number of shoots increased with increasing BAP concentrations, but this also resulted in a reduced rooting rate and root length. Plants acclimatized under 0 %, 35 % or 65 % of shading showed a dynamic maximum quantum yield of photosystem II (Fv/Fm), which initially decreased within the first seven days after the transfer to ex vitro conditions, but then increased until reaching stable values of 0.775 after 17 days. Additionally, the shading improved the plant survival rates, if compared to those under non-shaded conditions, which presented photoinhibition and photodamage symptoms.

RESUMO A contaminação por micro-organismos e oxidação dos explantes no estabelecimento in vitro de bambu são problemas recorrentes na sua micropropagação. Avaliaram-se os efeitos do biocida Plant Preservative Mixture (PPM®), do antibiótico casugamicina e da consistência do meio de cultura no estabelecimento in vitro de Bambusa vulgaris,Phyllostachys bambusoides e Dendrocalamus asper. A presença de PPM® no meio de cultura apresentou efeito significativo nas concentrações de 2 mL L-1 ou 4 mL L-1, assim como o meio de cultura líquido no aumento de plantas estabelecidas no outono. A casugamicina promoveu respostas variáveis para as três espécies, dependendo da estação do ano. Houve interação entre os fatores, demonstrando que maiores índices de plantas viáveis podem ser obtidos aliando-se diferentes estratégias para diminuir a oxidação e a contaminação. Para o estabelecimento in vitro de B. vulgaris,P. bambusoides e D. asper, é recomendada a adição de 2 mL L-1 ou 4 mL L-1 de PPM® ao meio de cultura líquido.

ABSTRACT The contamination by microorganisms and oxidation of explants in the in vitro establishment of bamboo are recurrent problems for its micropropagation. In the present study, effects of the biocide Plant Preservative Mixture (PPM™), the antibiotic kasugamycin and the consistency of the culture medium were evaluated in the in vitro establishment of Bambusa vulgaris,Phyllostachys bambusoides and Dendrocalamus asper. The presence of PPM™ in the culture medium had a significant effect using 2 mL L-1 or 4 mL L-1 concentrations, as well as in the liquid culture medium, increasing the plants established in the autumn. Kasugamycin promoted variable responses for all the three species, depending on the season. There was interaction among the factors, demonstrating that higher rates of viable plants can be obtained by combining different strategies to reduce the oxidation and contamination. For the in vitro establishment of B. vulgaris,P. bambusoides and D. asper, it is recommended to add 2 mL L-1 or 4 mL L-1 of PPM™ to the liquid culture medium.

RESUMO Foi desenvolvido um método eficiente de regeneração de plantas para Aspidosperma polyneuron a partir de culturas nodais de plântulas. Sementes maduras foram desinfestadas e os embriões germinaram em meio de cultura Woody Plant Medium (WPM). Segmentos nodais do epicótilo e hipocótilo retirados de plântulas de três semanas crescendo in vitro foram cultivados em meio WPM, suplementado com 6-benziladenina (BA) (2,5; 5,0 e 10 µM), isolado ou combinado com ácido indol-3-butírico (AIB) ou ácido α-naftalenoacético (ANA) (0,5 µM) durante o cultivo inicial e três subcultivos. Para indução de raízes, inicialmente, foram testados tratamentos-pulso de AIB (2,5; 5,0 ou 10 mM) por 15 minutos, seguidos de transferência para meio WPM, sem regulador de crescimento por cinco semanas. Plantas regeneradas foram transplantadas para bandejas contendo solo e substrato Plantmax® (3:1) e depois transferidas para sacos plásticos para aclimatização em casa de vegetação. Os explantes do hipocótilo apresentaram resultados superiores de regeneração de brotações e enraizamento, quando comparados com os do epicótilo. Os maiores números de brotações por explante (7-8) foram obtidos em meios de cultura, suplementados com 10 µM de BA, no terceiro subcultivo. Aadição de 0,5 µM de AIB ou ANA combinada com BAnão influenciou a regeneração de brotações. O tratamento-pulso de AIB (5-10 mM) por 15 minutos induziu 60% de enraizamento. As plantas regeneradas foram aclimatizadas e bem estabelecidas em casa de vegetação com 90% de sobrevivência, após três meses. Baseado nos resultados desse estudo, a micropropagação de Aspidosperma polyneuron a partir de plântulas cultivadas in vitro é viável e pode ser uma boa opção para conservação e propagação dessa espécie importante que está em extinção.

ABSTRACT In this study, an efficient method for regenerating plants from nodal cultures of seedlings was developed for Aspidosperma polyneuron. Mature seeds were surface-sterilized and embryos were germinated in Woody Plant Medium (WPM). Epicotyl and hypocotyl nodal segments, excised from 3-week-old in vitro-grown seedlings, were cultured in WPM medium supplemented with 6-benzyladenine (BA) (2.5, 5.0, and 10µM), alone or combined with indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA) (0.5 µM) for culture initiation and three subcultures. For root induction, IBA (2.5, 5.0, and 10 mM) pulse treatments (15 minutes) were initially applied, followed by transfer to growth regulator-free WPM for five weeks. Regenerated plants were first transplanted to trays containing soil and Plantmax® substrate (3:1) and later to polyethylene bags for acclimatization in a greenhouse. Hypocotyl explants exhibited superior shoot regeneration rate and rooting percentages compared with those of epicotyl explants. The highest numbers of shoots per explant (7−8) were obtained at the third subculture when using the culture medium supplemented with 10 µM BA. When combined with BA, the addition of 0.5 µM IBA or NAA did not affect the regeneration of shoots. An IBA pulse treatment of 5−10 mM for 15 minutes induced 60% of rooting. The regenerated plants were acclimatized and successfully established in a greenhouse, with a 90% survival rate observed after three months. Based on the results of this study, the micropropagation of Aspidosperma polyneuron from in vitro seedlings is feasible and could be a useful tool for the conservation and propagation of this important endangered species.

Protocolos de micropropagação de plantas possibilitam a produção de mudas isentas de pragas e doenças. No presente trabalho, objetivando a otimização da micropropagação do abacaxizeiro [Ananas comosus (L.) Merril], foram avaliados três fatores: 1) sistema de imersão (temporária por frascos duplos em erlenmeyers de 2 L ou permanente); 2) meio de cultura (suplementado, ou não, com Ácido Giberélico); 3) classe de altura de brotos. Os efeitos na taxa de regeneração (número de novos brotos para cada 1 g de massa fresca (MF) de microbrotos inoculada), na MF por broto, e na sobrevivência dos brotos foram avaliados. Interações significativas (p<0,05) foram observadas na taxa de regeneração, e efeitos simples (p<0,05) foram detectados na MF por broto. Todos os tratamentos resultaram em sobrevivência de mudas superior a 95% na aclimatização. Concluiu-se que a combinação do sistema de imersão temporária em frascos duplos e o meio de cultura isento de AG3 resultou na otimização do protocolo regenerativo. Esta combinação também demonstrou resposta positiva em um sistema de imersão temporária usual em biofábricas, que utiliza frascos duplos em garrafas PET de 5L.

Micropropagation based techniques enable the mass production of pest- and disease-free stock plants. Aiming at optimizing pineapple [Ananas comosus (L.) Merril] micropropagation, a complete randomized experiment with three factors was established: immersion system (2 L twin flasks temporary immersion and permanent immersion); culture medium (supplemented or not with GA3); and shoot height class. The effects on regeneration rate (number of new shoots per 1 g fresh mass (FM) of inoculated microshoots), FM per shoot, and shoot survival were evaluated. Significant interactions (p<0.05) were observed in the regeneration rate, and simple effects (p<0.05) were detected for FM per shoot. All treatments resulted in a survival rate higher than 95%. Altogether, it was found that 2 L twin flasks temporary immersion and MS culture medium free of GA3 allowed the optimization of the protocol used. This treatment was tested with positive response in a biofactory system that uses sterile plastic bottles (5 L).

A agrobiodiversidade é a parcela da biodiversidade constituída por um conjunto de organismos e ecossistemas que apresentam fortes relações com os seres humanos, podendo ser domesticados, semidomesticados, cultivados ou manejados pelo homem. Essa diversidade, conservada há séculos pelo uso (conservação in situ ou conservação on farm) pelos pequenos agricultores em todas as regiões do planeta, encontra-se ameaçada. De um lado, a agricultura industrial ou química através de suas práticas, ricas em insumos químicos, provoca muitas externalidades negativas, até recentemente pouco consideradas. De outro lado, as mudanças climáticas, igualmente, produzem efeitos adversos à diversidade genética. No entanto, as preocupações com a produção de alimentos de alta qualidade biológica em sistemas agrícolas sustentáveis remontam ainda o início do século XX. Dentre os vários sistemas agrícolas sustentáveis ou alternativos, a agroecologia apresenta vários atributos favoráveis: não faz uso de insumos químicos, é sustentável em todas as suas dimensões, faz uso de grande diversidade genética no cultivo, é socioeconômico-associada, estimula as relações de vizinhança e produz alimentos com alta qualidade biológica e nutricional. No âmbito da agroecologia e não da agricultura industrial ou química, há inúmeras oportunidades para a C&T desenvolver pesquisas participativas, contextualizadas, que podem empoderar tanto a agricultura familiar quanto as comunidades tradicionais no aperfeiçoamento dos processos e princípios agroecológicos utilizados. Assim, muitas das externalidades negativas atualmente inadmissíveis poderiam ser evitadas.

Agrobiodiversity is the portion of biodiversity consisting of a set of organisms and ecosystems that has strong relationships with humans, being domesticated, semi-domesticated, cultivated or managed by human beings. This diversity, which has been preserved by the practices (in situ on-farm conservation) of smallholder farmers in all regions of the world, is under threat. On one hand, industrial or chemical agriculture, because their practices are rich in external chemical inputs, causes many negative externalities that have not been considered much until recently. On the other hand, climate changes also produce adverse effects on the genetic diversity. However, since the beginning of the twentieth century concerns about the production of high quality organic foods in sustainable agricultural systems have been affirmed. Among several alternative or sustainable farming systems, agroecology comes with many favorable features: it makes no use of agrochemicals, it is environmentally sustainable, it makes use of high genetic diversity in cropping, it is socio-economically associated, it stimulates neighborhood relations, and it has food products of high biological quality. In the context of agroecology, but not in industrial or chemical agriculture, there are numerous opportunities for science and technology to develop participatory research, contextualized, that can empower family farming as well traditional communities in improving agroecological principles and processes. Thus, many of the currently unacceptable negative externalities could be avoided.

A Mata Atlântica é um hotspot de biodiversidade, e as bromélias são um grupo taxonômico bastante comum presente neste bioma. O objetivo deste estudo foi caracterizar os aspectos morfológicos e histoquímicos das sementes Vriesea friburgensis var paludosa L.B. Sm. As sementes são filiformes, com tegumento marrom e apêndices plumosos amarelados. Têm valores médios de 4,4 mm de comprimento, 0,5 mm de largura, e peso de 1.000 sementes de 683,8 mg. O tegumento apresenta compostos fenólicos. O amido é o principal componente de reserva no endosperma, porém ele também se apresenta em menores quantidades no embrião. O embrião ocupa um terço da semente e apresenta uma zona de constrição no eixo hipocotilo-radicula. Os resultados do presente trabalho contribuem na biologia e na delimitação taxonomia de V. friburgensis var. paludosa

The Brazilian Atlantic Forest is a biodiversity hotspot, and bromeliads are omnipresent in this biome. This study was aimed at characterizing both morphological and histochemical aspects of Vriesea friburgensis var. paludosa L.B. Sm. seeds. The seeds are filiform with a brown seed coat and yellowish-colored plumose appendages. Seeds are typically 4.4 mm long, 0.5 mm wide, and 1000 seeds weigh 683.8 mg. The seed coat presents phenolic compounds. Starch represents the main reserve compound in the endosperm; however, it is also present in smaller amounts in the embryo. The embryo occupies one-third of the seed and has a zone of constriction in the hypocotyl-radicle axis. The results of this work will contribute to the overall biology of V. friburgensis var. paludosa , as well as its taxonomic delimitation.

Angiosperm and gymnosperm plants evolved from a common ancestor about 300 million years ago. Apart from morphological and structural differences in embryogenesis and seed origin, a set of embryogenesis-regulating genes and the molecular mechanisms involved in embryo development seem to have been conserved alike in both taxa. Few studies have covered molecular aspects of embryogenesis in the Brazilian pine, the only economically important native conifer in Brazil. Thus eight embryogenesis-regulating genes, viz.,ARGONAUTE 1, CUP-SHAPED COTYLEDON 1, WUSCHEL-related WOX, S-LOCUS LECTIN PROTEIN KINASE, SCARECROW-like, VICILIN 7S, LEAFY COTYLEDON 1, and REVERSIBLE GLYCOSYLATED POLYPEPTIDE 1, were analyzed through semiquantitative RT-PCR during embryo development and germination. All the eight were found to be differentially expressed in the various developmental stages of zygotic embryos, seeds and seedling tissues. To our knowledge, this is the first report on embryogenesis-regulating gene expression in members of the Araucariaceae family, as well as in plants with recalcitrant seeds.

O quiwizeiro é uma espécie dióica, necessitando de polinização para a produção dos frutos. O objetivo deste trabalho foi avaliar a viabilidade in vitro e o tempo de conservação dos grãos de pólen de duas variedades de quiwi. Foram coletadas flores das variedades Matua e Tomuri. A germinação in vitro foi realizada em meio de cultura, contendo ágar (1 %), sacarose (0, 5, 10, 20 e 40 %) e ácido bórico (0 e 50 mg L-1 de H3BO3). Os grãos de pólen foram mantidos em BOD (25,0 ºC), geladeira (4,0 ºC), freezer (-18,0 ºC) e em N2 líquido (-196,0 ºC). As avaliações foram realizadas aos 0, 40, 120, 240 e 365 dias de conservação. O meio de cultura mais adequado foi suplementado com 12 % de sacarose e 50 mg L-1 de H3BO3. Os grãos de pólen podem ser conservados por curto período em geladeira ou freezer, e por um ano, sob criopreservação.

Kiwi is a dioecious plant species, requiring cross pollination for fruit production. The objective of this study was to evaluate the in vitro viability and shelf life of pollen grains of two kiwi varieties. Flowers of the cultivars Matua and Tomuri were collected and the pollen germinated in vitro, in culture medium containing agar (1 %), sucrose (0, 5, 10, 20 and 40 %) and boric acid (0 and 50 mg L-1 H3BO3). Pollen grains were stored in a BOD incubator (25.0 °C), refrigerator (4.0 °C), freezer (-18.0 °C) and in liquid N2 (-196.0 °C), and evaluated after 0, 40, 120, 240 and 365 days. The culture medium enriched with 12 % sucrose and 50 mg L-1 H3BO3 was the most suitable. Pollen grains can be stored for a short period in the refrigerator or freezer, and cryopreserved for at least one year.

A variedade Goethe é símbolo da vitivinicultura da região de Urussanga, sul do Estado de Santa Catarina, a qual, atualmente, busca a Indicação Geográfica da Uva e do Vinho Goethe. Para isto, um dos requisitos necessários é a identificação precisa do material genético. Os marcadores microssatélites constituem a ferramenta molecular mais utilizada para a identificação varietal de videira em todo o mundo e têm a capacidade de produzir um perfil genético único para cada material vitícola. O objetivo deste trabalho foi caracterizar duas seleções de uva 'Goethe', presentes no município de Urussanga, por meio de marcadores moleculares microssatélites, visando a atender aos requisitos de denominação de origem e indicação de procedência controlada. A extração do DNA genômico foi realizada a partir de folhas jovens e ramos de nove acessos de cada seleção de 'Goethe Classica' e 'Goethe Primo' provenientes de uma coleção pública e de oito coleções privadas da região de Urussanga. Dez loci microssatélites VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79, VVMD25, VVMD28, VVMD31 e VVMD32 foram genotipados através de eletroforese capilar. As análises realizadas mostraram que as duas variantes da uva 'Goethe' apresentaram um perfil molecular idêntico e único, isto é, representam a mesma variedade e sem nenhuma correspondência com variedades descritas anteriormente na literatura e nos bancos de dados consultados. As diferenças fenotípicas observadas provavelmente são devidas a mutações somáticas em regiões funcionais do genoma, fenômeno que dá origem aos clones em videira.

'Goethe' grape is a symbol of the viticulture of Urussanga region, South of Santa Catarina State, which is currently claiming the geographical indication of the grape and wine Goethe. Microsatellite markers are the biotechnological tool most used for molecular identification of grapevine varieties worldwide. These markers have the potential of generate unique DNA profiles for all the diverse grape genotypes. The aim of this work was to characterize two selections of the grapevine 'Goethe' from Urussanga, using microsatellite markers. Genomic DNA was extracted from young leaves and branches of nine accessions of each selection "Goethe Clássica and 'Goethe Primo' maintained at one public and eight private collections of Urussanga. Ten microsatellite loci VVS2, VVMD5, VVMD7, VVMD27, VrZAG62, VrZAG79, VVMD25, VVMD28, VVMD31 and VVMD32 were genotyped by capillary electrophoresis. Results showed that the two variants of 'Goethe' have an identical molecular profile which doesn't match any other DNA profile of grapevines previously described in literature or available in databases. The phenotypic differences observed are probably due to somatic mutations in functional regions of the genome, a phenomenon which leads to the creation of clones in grape.