Clonagem de quatro espécies de Annonaceae potenciais como porta-enxertos

O trabalho objetivou o estudo da capacidade de enraizamento de quatro espécies de Annonaceae potenciais como porta-enxertos (Annona glabra L., Annona montana Macfad, Rollinia emarginata e Rollinia mucosa Baill.) em três épocas do ano (verão, outono e inverno). Experimento conduzido em câmara de nebulização intermitente pertencente à UNESP/FCAV (21º15'22S e 48º18'58W), utilizando-se de estacas apicais enfolhadas em tratamento rápido (5 segundos) com ácido indolbutírico (IBA) (0; 1000; 3000; 5000 e 7000 mg.L-1), em esquema fatorial. Após 90 dias, avaliaram-se a porcentagem de estacas enraizadas, sobrevivência, comprimento e número de raízes. Como resultados, o IBA não incrementou a capacidade de enraizamento das espécies. A época do ano foi grande influente no sucesso de enraizamento, sendo o verão mais adequado. A. glabra e A. montana mostraram-se promissoras na propagação vegetativa via estaquia (clonagem). R. emarginata apresentou baixos valores para as características avaliadas, devendo ser intensificados experimentos na promoção do enraizamento em relação aos atuais valores obtidos. R. mucosa não apresentou resultados satisfatórios, ocorrendo necrose de tecido na base das estacas, devendo as causas serem melhor investigadas. O sucesso no enraizamento de Annonaceae é dependente da espécie, tendo aquelas do gênero Annona apresentado resultados superiores em relação às do gênero Rollinia.

Meios de cultura no desenvolvimento de ápices caulinares de mamoneira (Ricinus communis L.) in vitro

A pesquisa da composição do meio de cultura mais adequado à espécie vegetal e ao tipo de explante empregado é o fator de maior relevância da cultura de tecidos. O cultivo de ápice caulinar com recuperação da planta matriz é uma técnica de grande impacto para a propagação de plantas in vitro, regeneração de plantas livres de vírus, conservação de germoplasma e modificação genética. Objetivou-se, neste trabalho, avaliar composições do meio de cultivo para organogênese direta in vitro a partir de ápices caulinares pertencentes à população FCA-UNESP-PB de mamoneira (Ricinus communis L.), com vistas à propagação clonal de genótipos elite. Foram testadas quatro formulações: MS básico (T1), MS modificado 1 (T2), MS modificado 2 (T3) e WPM (T4), em delineamento experimental inteiramente casualizado, com 20 repetições em cada tratamento, sendo a repetição 1 ápice caulinar/frasco. O T3 apresentou-se superior e diferiu significativamente dos outros tratamentos apresentando 35% dos ápices caulinares diferenciados e desenvolvidos; seguiu-se o T2 com 10% e os tratamentos T1 e T4 não apresentaram diferenciação de tecidos. Os resultados permitiram concluir que os balanceamentos de sais minerais nos meios de cultura avaliados, especialmente a relação NO3 / NH4 e ausência de FeSO4.7H2O, indicaram grande influência no desenvolvimento de ápices caulinares de mamoneira.

Desenvolvimento in vitro de copo-de-leite: efeito das concentrações de sacarose e de ácido giberélico

Calla lily (Zantedeschia aethiopica) is appreciated as cut flower and for the composition of gardens. However, many pathogens affect this species. By the traditional method of propagation, some units of new seedlings can only be produced annually. Tissue culture allows fast large-scale clonal propagation and provides healthy uniform plants. During the in vitro process, type and concentration of growth regulator could affect the growth of seedlings. Thus, the aim of this work was to determine sucrose and GA(3) concentrations to increase the efficiency of the in vitro multiplication of calla lily. After 60 days, the length of the above ground part and the roots, the number of sprouts, roots and leaves, above ground part and root fresh weight of seedlings were evaluated. The experimental design was entirely randomized with four replications. It was necessary the addition of 60.5 g L-1 sucrose associated to 5 mg L-1 GA(3) to obtain hight sprouts number. For higher length of the above ground part the addition of 45.3 g L-1 sucrose and 10 mg L-1 GA(3) was enough. Better results in the root length and number of roots were observed only in the sucrose presence, in concentrations in the range of 51.13 - 56.5 g L-1.

Bud source, asepsis and benzylaminopurine (BAP) effect on yacon (Polymnia sonchifolia) micropropagation

The yacon (Polymnia sonchifolia) is used largely for the high fructan content of its tubers; consequently, it is a good alternative for diabetics. One of the more important restricting factors of the commercial production of yacon is its susceptibility to nematode attack. This, as well as germplasm bank maintenance, justifies the importance of in vitro propagation of this species. In this way, our work aimed to verify the best asepsis method for yacon for the in vitro establishment from the rhizophore and the axillary buds of the aerial parts, and the effect of benzylaminopurine (BAP) addition to the culture medium. The number of contaminated cultures, the occurrence of phenolic oxidation and the occurrence of a vitreous aspect, showed differences with bud source, immersion time for asepsis, and BAP use. The results contribute to establishing a yacon micro propagation procedure.

Micropropagação de Calendula officinalis L

Micropropagation of Calendula officinalis L. is usually propagated through seeds and therefore shows high diversity in flower size and colour, what causes quantitative and qualitative chemical variability. A micropropagation protocol was established for clonal propagation of this species to achieve homogeneous biomass, more appropriate for the production of phytotherapics. Explants harvested from capitula were the most appropriate for the micropropagation process. MS culture medium supplemented with 1.0 mgL-1 BAP and 6.0 gL-1 Phytagel™ enhanced shoot proliferation, while MS medium supplemented with 0.5 mgL-1 Kinetin and 6.0 gL-1 Phytagel™, increased shoot elongation. Plantlets (80%) cultured in MS/2 medium supplemented with 1.0 mgL-1 de IBA rooted.

Dinâmica espaço-temporal e quantificação dos danos e perdas da murcha de Ceratocystis em eucalipto na produção de carvão vegetal

Pós-graduação em Ciência Florestal - FCA

Gene expression during early somatic embryogenesis in Brazilian pine (Araucaria angustifolia (Bert) O. Ktze)

Brazilian pine (Araucaria angustifolia (Bert) O. Ktze) is the only native conifer species with economic importance in Brazil. Recently, due to intensive exploitation Brazilian pine was included in the official list of endangered Brazilian plants, under the "vulnerable" category. Biotechnology tools like somatic embryogenesis (SE) are potentially useful for mass clonal propagation and ex situ conservation strategies of commercial and endangered plant species. In spite of that, numerous obstacles still hamper the full application of SE technology for a wider range of species, including Brazilian pine. To enhance somatic embryogenesis in Brazilian pine and to gain a better understanding of the molecular events associated with somatic embryo development, we analyzed the steady-state transcript levels of genes known to regulate somatic embryogenesis using semiquantitative reverse transcription polymerase chain reaction (sqRT-PCR). These genes included Argonaute (AaAGO), Cup-shaped cotyledon1 (AaCUC), wushel-related WOX (AaWOX), a S-locus lectin protein kinase (AaLecK), Scarecrow- like (AaSCR), Vicilin 7S (AaVIC), Leafy Cotyledon 1 (AaLEC), and a Reversible glycosylated polypeptide (AaRGP). Expression patterns of these selected genes were investigated in embryogenic cultures undergoing different stages of embryogenesis, and all the way to maturation. Up-regulation of AaAGO, AaCUC, AaWOX, AaLecK, and AaVIC was observed during transition of somatic embryos from stage I to stage II. During the maintenance phase of somatic embryogenesis, expression of AaAGO and AaSCR, but not AaRPG and AaLEC genes was influenced by presence/ absence of plant growth regulators, both auxins and cytokinins. The results presented here provide new insights on the molecular mechanisms responsible for somatic embryo formation, and how selected genes may be used as molecular markers for Brazilian pine embryogenesis.

Growth data from a field trial of Quercus suber plants regenerated from selected trees and their half-sib progenies by somatic embryogenesis

The development of reliable clonal propagation technologies is a requisite for performing Multi-Varietal Forestry (MVF). Somatic embryogenesis is considered the tissue culture based method more suitable for operational breeding of forest trees. Vegetative propagation is very difficult when tissues are taken from mature donors, making clonal propagation of selected trees almost impossible. We have been able to induce somatic embryogenesis in leaves taken from mature oak trees, including cork oak (Quercus suber). This important species of the Mediterranean ecosystem produces cork regularly, conferring to this species a significant economic value. In a previous paper we reported the establishment of a field trial to compare the growth of plants of somatic origin vs zygotic origin, and somatic plants from mature trees vs somatic plants from juvenile seedlings. For that purpose somatic seedlings were regenerated from five selected cork oak trees and from young plants of their half-sib progenies by somatic embryogenesis. They were planted in the field together with acorn-derived plants of the same families. After the first growth period, seedlings of zygotic origin doubled the height of somatic seedlings, showing somatic plants of adult and juvenile origin similar growth. Here we provide data on height and diameter increases after two additional growth periods. In the second one, growth parameters of zygotic seedlings were also significantly higher than those of somatic ones, but there were not significant differences in height increase between seedlings and somatic plants of mature origin. In the third growth period, height and diameter increases of somatic seedlings cloned from the selected trees did not differ from those of zygotic seedlings, which were still higher than data from plants obtained from somatic embryos from the sexual progeny. Therefore, somatic seedlings from mature origin seem not to be influenced by a possible ageing effect, and plants from somatic embryos tend to minimize the initial advantage of plants from acorns

Obtención de clones de alcornoque (Quercus suber L.) mediante embriogénesis somática: aplicación de medios líquidos y biorreactores en la sincronización de los procesos de desarrollo y maduración de los embriones

El alcornoque tiene un gran valor ambiental, como integrante de los ecosistemas forestales mediterráneos, e interés comercial por el valor de la bellota (alimentación del cerdo ibérico), el carbón, la madera y sobre todo por las aplicaciones industriales del corcho. Las posibilidades de mejora genética del alcornoque, como las de otras especies forestales, están limitadas por sus largos ciclos reproductivos y porque su propagación vegetativa mediante estaquillado solo es posible en estados muy juveniles. Por ello este sistema de propagación tiene muy poca, o ninguna, utilidad práctica en la mejora genética. La embriogénesis somática es la vía más apropiada para la clonación de muchas especies forestales y ha hecho posible el desarrollo a gran escala de plantaciones multivarietales de coníferas. En alcornoque es posible la regeneración completa de árboles adultos mediante embriogénesis somática. Con los protocolos actuales (en medio semisólido), los embriones se generan formando acúmulos y en la fase de multiplicación conviven embriones en distintos estados de desarrollo. Es un sistema asincrónico, con baja eficacia para la propagación en masa, que no elimina completamente las dificultades para el desarrollo de programas de mejora genética del alcornoque. En otras especies la utilización de medios líquidos ha mejorado: la sincronización, productividad de los cultivos, el manejo y reducido los costes de producción. Por ello el desarrollo de suspensiones embriogénicas de alcornoque se plantea como una vía para aumentar la eficacia de la propagación clonal a gran escala. En la presente tesis se desarrollan cultivos embriogénicos de alcornoque en medio líquido. El capítulo 3 aborda el establecimiento y mantenimiento de suspensiones, el capítulo 4 el desarrollo de una fase de proliferación en medio líquido y el capítulo 5 la utilización de sistemas de cultivo en medio líquido, estacionarios y de inmersión temporal, como vía para favorecer la maduración de los embriones somáticos. Para iniciar los cultivos en medio líquido se emplearon agregados de embriones tomados de la fase de proliferación en medio semisólido. Cuando estos agregados se inocularon directamente en medio líquido no se logró el establecimiento de las suspensiones. El establecimiento se consiguió empleando como inóculo las células y Resumen pequeños agregados embriogénicos, de tamaño comprendido entre 41 y 800 μm, desprendidas por agitación breve de los agregados de embriones. El mantenimiento se logró inoculando en baja densidad masas embriogénicas compactas de tamaño comprendido entre 0,8 y 1,2 mm. Estas suspensiones, muy heterogéneas, mantuvieron su capacidad de proliferación y de regeneración de embriones al menos durante diez subcultivos consecutivos. El protocolo de iniciación y mantenimiento, desarrollado inicialmente con un solo genotipo, fue eficaz cuando se probó sobre otros 11 genotipos de alcornoque. En la fase de proliferación se ensayaron tres tipos de envase y tres velocidades de agitación. La combinación envase × velocidad determinó el intercambio gaseoso, la disponibilidad de oxígeno y el estrés hidrodinámico. Los agregados embriogénicos de alcornoque crecieron incluso en condiciones de hipoxia no siendo la disponibilidad de oxígeno un factor limitante del crecimiento para tasas de trasferencia de oxígeno comprendidas entre 0,11 h-1 y 1,47 h-1. Por otra parte la producción de biomasa creció con el estrés hidrodinámico para valores de índice de cizalladura inferiores a 5 x 10-3 cm min-1. La mayor producción de biomasa se obtuvo con matraces Erlenmeyer de 100 ml y alta velocidad de agitación (160 rpm) mientras que la diferenciación de embriones se vio favorecida por bajas velocidades de agitación (60 rpm) asociadas con bajas disponibilidades de oxígeno. La posibilidad de madurar embriones de alcornoque en medio líquido se estudió utilizando sistemas de inmersión permanente y sistemas de inmersión temporal. En inmersión permanente no se diferenciaron embriones cotiledonares (posiblemente por hiperhidricidad). Los sistemas de inmersión temporal permitieron obtener embriones maduros en estado cotiledonar y capaces de regenerar plantas in vitro. Concentraciones de sacarosa superiores a 60 g l-1 y frecuencias de inmersión iguales o inferiores a una diaria, tuvieron efectos negativos para el desarrollo de los embriones somáticos. En los sistemas de inmersión temporal los parámetros físico-químicos del medio de cultivo se mantuvieron estables y no se observó ninguna limitación de nutrientes. No obstante, estos sistemas se vieron afectados por la evaporación que generó el flujo de aire necesario para desplazar el líquido en cada periodo de inmersión. Abstract ABSTRACT Cork oak is one of the most important tree species of the Mediterranean ecosystem. Besides its high environmental value has a great economic interest due to the sustainable production of acorns (to feed the Iberian pig) charcoal, timber and cork, which is a renewable natural product with various technological applications. As happens with other forest species, cork oak genetic improvement programs are limited by their long life cycles and because vegetative propagation by cuttings it´s only possible in very juvenile plants. Hence this propagation system is useless or has little practical use for breeding cork oak. Plant regeneration by somatic embryogenesis is the most suitable way for cloning many forest species, and it is the enabling technology which has allowed the establishment of large-scale conifer multi-varietal plantations. Clonal plant regeneration of mature cork oak trees can be achieved through somatic embryogenesis. Somatic embryos at different stages of development and forming clusters are produced during the multiplication phase with current protocols (using semisolid medium). This is an asynchronous low-efficient process not suitable for mass propagation, and therefore it does not solve the difficulties presented by cork oak breeding programs. Culture in liquid medium has been used with other species to improve: synchronization, yield, handling, and to reduce production costs. Thus the development of cork oak embryogenic suspension cultures is envisaged as a way to increase the efficiency of large scale clonal propagation. The thesis herein develops cork oak embryogenic cultures in liquid medium. In chapter 3 establishment and maintenance of suspension cultures are developed, chapter 4 studies proliferation phase in liquid medium and chapter 5 considers the use of different systems of culture in liquid medium, both stationary and temporary immersion, as a way to promote somatic embryos maturation. Clusters of embryos taken from proliferating cultures on semisolid medium were used to initiate the cultures in liquid medium. When these clusters were inoculated directly in liquid medium establishment of suspension cultures was not executed. However using, as initial inoculum, cells and cell aggregates with a size between 41 and 800 μm detached from these clusters of embryos, subjected to a brief shaking, suspension cultures could be established. Suspension maintenance was achieved by inoculating compact embryogenic Abstract clumps with a size between 0.8 and 1.2 mm at low density. The suspension cultures, very heterogeneous, retained both their proliferation and embryo regeneration capacity for at least ten consecutive subcultures. The initiation and maintenance protocol, initially developed with a single genotype, was effective when tested on 11 additional genotypes of cork oak. In proliferation phase three types of vessels and three different levels of agitation were assayed. The combination vessel × orbiting speed determined gas exchange, oxygen availability and hydrodynamic stress. Cork oak embryogenic aggregates grew even under hypoxia conditions; oxygen availability at transfer rates between 0.11 and 1.47 h-1 was not a limiting factor for growth. Furthermore the biomass production was increased with hydrodynamic stress when shear rate values were of less than 5 x 10-3 cm min-1. The highest biomass production was obtained with 100 ml Erlenmeyer flask and high stirring speed (160 rpm) while the differentiation of embryos was favored by low agitation speeds (60 rpm) associated with low oxygen availability. The possibility to mature cork oak somatic embryos in liquid medium was studied using both permanent immersion systems and temporary immersion systems. Cotyledonary embryos did not differentiate in permanent immersion conditions (probably due to hyperhydricity). Temporary immersion systems allowed obtaining mature cotyledonary embryos, which were able to regenerate plants in vitro. Sucrose concentrations above 60 g l-1 and immersion frequencies equal to or lower than one each 24 h had negative effects on somatic embryo development. Physicochemical parameters of the culture medium in temporary immersion systems were stable and showed no limitation of nutrients. However, these systems were affected by the evaporation generated by the airflow necessary to relocate the medium at each immersion period.

Genetic instability of cancer cells is proportional to their degree of aneuploidy

Genetic and phenotypic instability are hallmarks of cancer cells, but their cause is not clear. The leading hypothesis suggests that a poorly defined gene mutation generates genetic instability and that some of many subsequent mutations then cause cancer. Here we investigate the hypothesis that genetic instability of cancer cells is caused by aneuploidy, an abnormal balance of chromosomes. Because symmetrical segregation of chromosomes depends on exactly two copies of mitosis genes, aneuploidy involving chromosomes with mitosis genes will destabilize the karyotype. The hypothesis predicts that the degree of genetic instability should be proportional to the degree of aneuploidy. Thus it should be difficult, if not impossible, to maintain the particular karyotype of a highly aneuploid cancer cell on clonal propagation. This prediction was confirmed with clonal cultures of chemically transformed, aneuploid Chinese hamster embryo cells. It was found that the higher the ploidy factor of a clone, the more unstable was its karyotype. The ploidy factor is the quotient of the modal chromosome number divided by the normal number of the species. Transformed Chinese hamster embryo cells with a ploidy factor of 1.7 were estimated to change their karyotype at a rate of about 3% per generation, compared with 1.8% for cells with a ploidy factor of 0.95. Because the background noise of karyotyping is relatively high, the cells with low ploidy factor may be more stable than our method suggests. The karyotype instability of human colon cancer cell lines, recently analyzed by Lengnauer et al. [Lengnauer, C., Kinzler, K. W. & Vogelstein, B. (1997) Nature (London) 386, 623–627], also corresponds exactly to their degree of aneuploidy. We conclude that aneuploidy is sufficient to explain genetic instability and the resulting karyotypic and phenotypic heterogeneity of cancer cells, independent of gene mutation. Because aneuploidy has also been proposed to cause cancer, our hypothesis offers a common, unique mechanism of altering and simultaneously destabilizing normal cellular phenotypes.

Modeling Nymphoides architecture: A morphological analysis of Nymphoides aquatica (Menyanthaceae)

• Premise of the study: Species in the aquatic genus Nymphoides have inflorescences that appear to arise from the petioles of floating leaves. The inflorescence-floating leaf complex can produce vegetative propagules and/or additional inflorescences and leaves. We analyzed the morphology of N. aquatica to determine how this complex relates to whole plant architecture and whether whole plant growth is sympodial or monopodial. • Methods: We used dissections, measurements, and microscopic observations of field-collected plants and plants cultivated for 2 years in outdoor tanks in south Florida, USA. • Key results: Nymphoides aquatica had a submerged plagiotropic rhizome that produced floating leaves in an alternate/spiral phyllotaxy. Rhizomes were composed of successive sympodial units that varied in the number of leaves produced before the apex terminated. The basic sympodial unit had a prophyll that subtended a renewal-shoot bud, a short-petioled leaf (SPL) with floating lamina, and an inflorescence; the SPL axillary bud expanded as a vegetative propagule. Plants produced either successive basic sympodial units or expanded sympodia that intercalated long-petioled leaves between the prophyll and the SPL. • Conclusions: Nymphoides aquatica grows sympodially, forming a rhizome composed of successive basic sympodia and expanded sympodial units. Variations on these types of sympodial growth help explain the branching patterns and leaf morphologies described for other Nymphoides species. Monitoring how these two sympodial phases are affected by water depth provides an ecologically meaningful way to assess N. aquatica’s responses to altered hydrology.

MULTIPLICAÇÃO E ALONGAMENTO IN VITRO DE CLONES HÍBRIDOS DE Eucalyptus globulus

Os híbridos de Eucalyptus globulus representam uma excelente alternativa para o setor de celulose e papel, em razão dos ganhos em qualidade da madeira para a fabricação de celulose. Entretanto, estes híbridos têm apresentado recalcitrância ao enraizamento adventício. Assim, a micropropagação é apontada como a técnica para o rejuvenescimento desses híbridos adultos, viabilizando a propagação clonal dos mesmos. O presente trabalho avaliou o cultivo in vitro de três clones de Eucalyptus grandis x Eucalyptus globulus e de três clones de Eucalyptus urophylla x Eucalyptus globulus, em relação à multiplicação in vitro, no meio MS suplementado com 0,5 mg L-1 de BAP e 0,01 mg L-1 de ANA, bem como o efeito das concentrações de 0,25; 0,50; 0,75 e 1,0 mg L-1 de AIB e dos meios de cultura MS e JADS no alongamento in vitro das brotações. Os clones diferiram quanto à multiplicação in vitro das brotações e apresentaram uma taxa de multiplicação média dos clones de 3,0 tufos de brotações em cada subcultivo, ao longo dos 25 subcultivos realizados. No alongamento in vitro, os clones diferiram quanto às concentrações de AIB adequadas para provocar o alongamento, bem como em relação aos meios de cultura MS e JADS. O intervalo médio entre 0,40 e 0,80 mg L-1 de AIB proporcionou o maior número e comprimento das brotações alongadas in vitro e com maior vigor.

VEGETATIVE RESCUE AND CUTTINGS PROPAGATION OF Araucaria angustifolia (Bertol.) Kuntze

Brazilian pine or araucaria (Araucaria angustifolia) is a coniferous tree with great economic, social and environmental importance in southern Brazil, being exploited for both wood production and for its edible pine nuts. However, no efficient cloning techniques are available and, therefore, the purpose of this study was to evaluate the effectiveness of vegetative rescue methods for cuttings propagation of the species. Shoots/cuttings were generated in two ways: 26 years old trees underwent coppicing and 20 years old trees had the primary branches on the upper third of crown pruned at 2, 20 and 50 cm from the main trunk. Orthotropic shoots were rooted after application of indole-3-butyric acid (IBA) at 0, 2, 4 and 6 g.L-1. Coppicing produced 47 cuttings per plant with 90% orthotropic shoots, while pruning resulted in 182 cuttings per plant with 44% orthotropic shoots. Rooting success indexes were low with no influence of IBA, although they are slightly superior to the ones available in the literature for the species, ranging from 12 to 30% for the coppice shoots and from 0 to 28% for the branches shoots. We conclude that both vegetative rescue techniques are viable and have potentially important applications. Coppicing is recommended for the propagation aiming the production of wood, while shoots derived from the side branches of the crown are more appropriate for seeds orchards formation.

ROOTING MINI-CUTTINGS OF Paulownia fortunei var. mikado DERIVED FROM CLONAL MINI-GARDEN1

ABSTRACTWe aimed to evaluate the technical efficiency of mini-cuttings technique on vegetative propagation of Paulownia fortunei (Seem.) Hemsl. var. Mikado, as well as the possible existence of anatomical barriers to its rooting. Therefore, plants originated from cuttings formed the mini-stumps and, consequently the clonal mini-garden, which was conducted in semi-hydroponic system. We evaluated the survival of mini-stumps and sprouts production for five successive collects, the percentage of mini-cuttings rooting and their anatomical description. The mini-cuttings were prepared with about 6 to 8 cm in length and two leaves reduced by about 50% in the upper third, being remained an area of, approximately 78 cm2 (10 cm diameter). The mini-cuttings were placed in tubes of 53 cm3, with substrate formed with fine vermiculite and carbonized rice hulls (1:1 v/v) and rooted in acclimatized greenhouse. After 30 days we evaluated the percentage of rooted mini-cuttings, radicial vigor (number and length of roots / mini-cutting), callus formation, emission of new shoots and maintenance of the original leaves. The mini-stumps showed 100% survival after five collects and an average production of 76-114 mini-cuttings/m2/month and rooting ranged from 70 to 90%. Mini-cuttings technique is efficient in to propagate adult propagules of the species and there are not anatomical barriers preventing roots emission.

Vegetative propagation strategies of four rupestrian species of Leiothrix (Eriocaulaceae)

Leiothrix is endemic of South America and includes 37 species, 25 of which occur in the state of Minas Gerais. Nineteen of those occur in the "Serra do Cipó", a mountain chain, located in the southern portion of the Espinhaço mountain range. This study examines vegetative propagation strategies of four species of Leiothrix, endemic to the Minas Gerais portion of the Espinhaço mountain range. For each species we established permanent plots, where we marked 30 to 51 rosettes or clones, and then took morphological and phenological measurements. Leiothrix crassifolia (Bong.) Ruhland and L. curvifolia var. lanuginosa (Bong.) Ruhland are rhizomatous, forming compact clones. Leiothrix vivipara (Bong.) Ruhland does not produce rhizomes, but is pseudoviviparous, i.e., produces numerous ramets originating from inflorescences. These ramets are formed precociously, and the flower heads do not touch the ground. In Leiothrix spiralis (Bong.) Ruhland both of these strategies are seen: it is both rhizomatous and pseudoviviparous. In this species, the ramets are formed late, only after the flower head has touched the ground. One of the typical conditions of the rupestrian grasslands is soil water shortage in some periods of the year and nutrient scarcity all year round. These conditions might have created an ideal ecological scenario for the evolution of both pseudovivipary and rhizomatous clonal growth in Leiothrix.