Caracterização isoenzimática de oito espécies do gênero Trifolium L., ocorrentes no Rio Grande do Sul

O presente estudo tem como objetivos: contribuir para o desenvolvimento de oito espécies do gênero Trifolium L., Trifolium incamatum L., Trifolium polymorphum Poir, Trifolium pratense L., Trifolium repens L., trifolium resupinatum L., Trifolium riograndense Burkart, Trifolium subterraneum L. e Trifolium vesiculosum Savi, através da caracterização da variabilidade isoenzimática presente em quatro sistemas enzimáticos, Fosfoglicoisomerase (PGI), Esterase (EST), Malato desidrogenase (MDH) e Superóxido dismutase (SOD) e fornecer subsídios para futuros trabalhos de melhoramento de plantas forrageiras.

Avaliação e melhoramento genético de trevo vermelho (Trifolium pratense L.) em duas regiões fisiográficas do Rio Grande do Sul

O trevo vermelho (Trifolium pratense L.) é uma leguminosa forrageira muito importante no Rio Grande do Sul, constituindo uma boa alternativa dentre as espécies forrageiras de inverno. Entretanto, como não existem cultivares desenvolvidas para as condições ambientais do Estado, a produção e, principalmente, a persistência dessas pastagens são comprometidas. Por isso o Departamento de Plantas Forrageiras e Agrometeorologia da UFRGS conduz um programa de melhoramento genético da espécie, buscando o desenvolvimento de populações melhor adaptadas às condições locais. O objetivo do trabalho foi avaliar, em dois ambientes contrastantes, Eldorado do Sul e Veranópolis, três populações de trevo vermelho selecionadas pelo referido programa, comparando-as com uma cultivar padrão. Em Eldorado do Sul foram implantados dois experimentos, um em 1999 e outro em 2000, enquanto em Veranópolis foi instalado apenas um, em 2000. Quanto à produção de matéria seca, os resultados variaram conforme o ano e o local. De modo geral, a cultivar padrão foi mais produtiva no primeiro corte, enquanto as populações selecionadas produziram mais forragem no final das estações de crescimento. Somente em Veranópolis o experimento foi avaliado por mais de um ano, com as populações selecionadas apresentando maior estabilidade produtiva que a cultivar padrão e produzindo mais forragem na segunda estação de crescimento. Em geral, as populações selecionadas também apresentaram maior persistência que o padrão. Ambos os locais mostraram-se favoráveis à avaliação e seleção para persistência do trevo vermelho, porém Veranópolis foi o mais adequado para a avaliação da produção de matéria seca.

Caracterização morfológica e molecular da coleção básica de trevo-branco (Trifolium repens L.)

O objetivo deste trabalhofoi avaliarcaracterísticas moleculares e morfológicasde 79 acessos pertencentes à coleção básica de trevo-branco obtida do Departamento de Agricultura dos Estados Unidos(USDA), a fim de caracterizar a variabilidade genética existente.

Seleção para aumento da produção de gametas não reduzidos e poliploidização sexual em trevo vermelho (Trifolium pratense L.)

Trevo vermellho (Trifolium pratense L.), é uma leguminosa forrageira de excelente qualidade, mas não persistente no Rio Grande do Sul. Plantas com maior variabilidade genética podem tornar esta espécie mais estável e produtiva. Poliplóides sexuais ocorrem em populações naturais e apresentam uma ampla base genética. Os objetivos deste trabalho foram: no Experimento 1, obter plantas poliplóides sexuais através de cruzamentos unilaterais e verificar a fertilidade desta descendência. No Experimento 2, cruzamentos bilaterais visaram aumentar a produção de gametas não reduzidos (2n) em plantas diplóides em três ciclos de seleção. No Experimento 1, a geração parental foi composta por tetraplóides somáticos e plantas diplóides da cv. Quiñiqueli, selecionadas por produzirem de 1 até 9,07% de gametas 2n. Em quatro gerações, a presença de indivíduos férteis triplóides indicou que estes podem ser utilizados como uma ponte para a produção de plantas tetraplóides. No Experimento 2, no primeiro ciclo, foram selecionadas plantas das cultivares Quiñiqueli, Redland e Keenland, que produziram no mínimo 1% de pólen gigante. No segundo e terceiro ciclos, plantas com no mínimo 2% e 3% de produção de pólen 2n foram selecionadas, respectivamente. A porcentagem de produção aumentou de 1,67% na população original para 8,97% na última geração de plantas selecionadas. O ganho de seleção foi de 437,12%, com diferencial de seleção de 7,3%. Os dados comprovaram que os métodos de detecção de grãos de gametas 2n e de seleção são eficientes, tornando possível obter plantas poliplóides sexuais em etapas avançadas, a partir de populações selecionadas de trevo vermelho.

Efeito de sistemas de pastejo sobre o desempenho e o comportamento de suínos em crescimento e terminação, mantidos em pastagem de trevo-branco (Trifolium repens L.)

O presente trabalho foi conduzido com o objetivo de avaliar o efeito de diferentes sistemas de pastejo (contínuo, alternado e rotativo) sobre o desempenho e o comportamento de suínos, assim como, sobre a disponibilidade e a composição química do trevo-branco (Trifolium repens L.). Foram utilizados 48 suínos machos, castrados, com peso médio inicial de 27 kg, sendo doze mantidos em sistema confinado, os quais foram utilizados como tratamento testemunha. Os animais foram distribuídos em um delineamento de blocos casualizados, com três tratamentos e três repetições. Não foram observadas diferenças significativas (P>0,05) entre os sistemas de pastejo nas características de desempenho e de disponibilidade. No entanto, foram observadas diferenças significativas (P<0,05) da composição química da pastagem entre os sistemas de pastejo estudados. Verificou-se que os suínos em sistemas de pastejo contínuo, alternado e rotativo consumiram de 13% a 17% a menos de ração (P<0,05) e apresentaram menores ganhos de peso (P<0,05) e espessura de toucinho (P<0,05) do que os suínos criados em confinamento. Não foram observadas diferenças significativas (P>0,05) na conversão alimentar entre os suínos mantidos em confinamento e sobre pastagem. Deitar, beber água, ingerir ração, pastejar e caminhar foram as atividades que apresentaram maiores freqüências de ocorrência, independente dos sistemas de pastejo estudados. Houve efeito significativo dos sistemas de pastejo sobre a atividade de ingerir ração (P=0,0186) e interação de sistema x dia de observação para as atividades de deitar (P=0,0345) e pastejar (P=0,0678).

Avaliação do potencial de produção de sementes de acessos de trevo branco (Trifolium repens L)

A produção de sementes de qualidade é um dos pontos chave na difusão do trevo branco em nosso meio. Esta leguminosa é uma espécie de boa produção de sementes, entretanto, como as demais espécies de Trifolium, é conhecida por flutuações no rendimento de sementes. Com o objetivo de avaliar o potencial de produção de sementes de 27 acessos da coleção básica de trevo branco conduziu-se o presente trabalho na região da Depressão Central do RS, EEA/UFRGS, Eldorado do Sul, no período de outubro de 2003 a março de 2004. As variáveis estudadas através de amostragens semanais realizadas entre 06/11 e 10/03 foram: número de inflorescências por planta, número de flores por inflorescência, número de inflorescências maduras, número de legumes maduros por inflorescência, peso de 1000 sementes e rendimento de sementes por planta. Foram separados três grupos distintos quanto à produção de sementes. O grupo superior com o acesso 53 (Peru), outro intermediário com os acessos 2 (Israel), 3 (EUA), 7 (Etiópia), 13 (Espanha), 19 (Costa Rica), 20 (Uruguai), 28 (Austrália), 33 (Irlanda), 50 (África do Sul), 58 (Índia), 64 (Suíça), 68 (Jamaica) e 75 (Uruguai) e um grupo inferior com os acessos 15 (Nova Zelândia), 22 (Hungria), 23 (Bélgica), 29 (França), 31 (EUA), 38 (Irã), 54 (Nova Zelândia), 59 (Líbano), 79 (Nova Zelândia) e 80 (EUA). O número de inflorescências, número de inflorescências maduras e peso de 1000 sementes apresentaram correlação positiva e altamente significativa em relação ao rendimento de sementes. O acesso 20 apresentou superioridade no número de inflorescências por planta e número de inflorescências maduras por planta, no entanto, o acesso 53 foi o de maior potencial de produção de sementes, em função de sua superioridade no peso de 1000 sementes. Os acessos 27 (Canadá), 65 (Dinamarca) e 73 (Polônia) não produziram sementes nas condições locais.

Host plant recognition by the root feeding clover weevil, Sitona lepidus (Coleoptera : Curculionidae)

This study investigated the ability of neonatal larvae of the root-feeding weevil, Sitona lepidus Gyllenhal, to locate white clover Trifolium repens L. (Fabaceae) roots growing in soil and to distinguish them from the roots of other species of clover and a co-occurring grass species. Choice experiments used a combination of invasive techniques and the novel technique of high resolution X-ray microtomography to non-invasively track larval movement in the soil towards plant roots. Burrowing distances towards roots of different plant species were also examined. Newly hatched S. lepidus recognized T. repens roots and moved preferentially towards them when given a choice of roots of subterranean clover, Trifolium subterraneum L. (Fabaceae), strawberry clover Trifolium fragiferum L. (Fabaceae), or perennial ryegrass Lolium perenne L. (Poaceae). Larvae recognized T. repens roots, whether released in groups of five or singly, when released 25 mm (meso-scale recognition) or 60 mm (macro-scale recognition) away from plant roots. There was no statistically significant difference in movement rates of larvae.

Commensalism in an agroecosystem: hydraulic redistribution by deep-rooted legumes improves survival of a droughted shallow-rooted legume companion

We investigated commensalism of water use among annual shallow-rooted and perennial deep-rooted pasture legumes by examining the effect of hydraulic lift by Cullen pallidum (N.T.Burb.) J.W.Grimes and Medicago sativa on growth, survival and nutrient uptake of Trifolium subterraneum L. A vertically split-root design allowed separate control of soil water in top and bottom soil. Thirty-five days after watering ceased in the top tube, but soil remained at field capacity in the bottom tube, an increase in shallow soil water content by hydraulic lift was 5.6 and 5.9 g kg−1 soil overnight for C. pallidum and M. sativa, respectively. Trifolium subterraneum in this treatment maintained higher leaf water potentials (with M. sativa) or exhibited a slower decline (with C. pallidum) than without companion perennial plants; and shoot biomass of T. subterraneum was 56% (with C. pallidum) and 67% (with M. sativa) of that when both top and bottom tubes were at field capacity. Uptake of rubidium (a potassium analog) and phosphorus by T. subterraneum was not facilitated by hydraulic lift. Interestingly, phosphorus content was threefold greater, and shoot biomass 1.5–3.3-fold greater when T. subterraneum was interplanted with C. pallidum compared with M. sativa, although dry weight of C. pallidum was much greater than that of M. sativa. This study showed that interplanting with deep-rooted perennial legumes has benefited the survival of T. subterraneum.

Nitrogen transfer within and between plants through common mycorrhizal networks (CMNs)

Mycorthizae play a critical role in nutrient capture from soils. Arbuscular mycorrhizae (AM) and ectomycorrhizae (EM) are the most important mycorrhizae in agricultural and natural ecosystems. AM and EM fungi use inorganic NH4+ and NO3-, and most EM fungi are capable of using organic nitrogen. The heavier stable isotope N-15 is discriminated against during biogeochemical and biochemical processes. Differences in N-15 (atom%) or delta(15)N (parts per thousand) provide nitrogen movement information in an experimental system. A range of 20 to 50% of one-way N-transfer has been observed from legumes to nonlegumes. Mycorrhizal fungal mycelia can extend from one plant's roots to another plant's roots to form common mycorrhizal networks (CMNs). Individual species, genera, even families of plants can be interconnected by CMNs. They are capable of facilitating nutrient uptake and flux. Nutrients such as carbon, nitrogen and phosphorus and other elements may then move via either AM or EM networks from plant to plant. Both N-15 labeling and N-15 natural abundance techniques have been employed to trace N movement between plants interconnected by AM or EM networks. Fine mesh (25similar to45 mum) has been used to separate root systems and allow only hyphal penetration and linkages but no root contact between plants. In many studies, nitrogen from N-2-fixing mycorrhizal plants transferred to non-N-2-fixing mycorrhizal plants (one-way N-transfer). In a few studies, N is also transferred from non-N-2-fixing mycorrhizal plants to N-2-fixing mycorrhizal plants (two-way N-transfer). There is controversy about whether N-transfer is direct through CMNs, or indirect through the soil. The lack of convincing data underlines the need for creative, careful experimental manipulations. Nitrogen is crucial to productivity in most terrestrial ecosystems, and there are potential benefits of management in soil-plant systems to enhance N-transfer. Thus, two-way N-transfer warrants further investigation with many species and under field conditions.

Reciprocal N ((NH4+)-N-15 or (NO3-)-N-15) transfer between nonN(2)-fixing Eucalyptus maculata and N-2-fixing Casuarina cunninghamiana linked by the ectomycorrhizal fungus Pisolithus sp.

Two-way N transfers mediated by Pisolithus sp. were examined by excluding root contact and supplying (NH4+)-N-15 or (NO3-)-N-15 to 6-month-old Eucalyptus maculata or Casuarina cunninghamiana grown in two-chambered-pots separated by 37 m screens. Mycorrhizal colonization was 35% in Eucalyptus and 66% in Casuarina (c. 29% N-2-fixation). Using an environmental scanning electron microscope, living hyphae were observed to interconnect Eucalyptus and Casuarina. Biomass and N accumulation was greatest in nodulated mycorrhizal Casuarina/mycorrhizal Eucalyptus pairs, less in nonnodulated mycorrhizal Casuarina/mycorrhizal Eucalyptus pairs, and least in nonnodulated nonmycorrhizal Casuarina/nonmycorrhizal Eucalyptus pairs. In nonnodulated mycorrhizal pairs, N transfers to Eucalyptus or to Casuarina were similar (2.4-4.1 mg per plant in either direction) and were 2.6-4.0 times greater than in nonnodulated nonmycorrhizal pairs. In nodulated mycorrhizal pairs, N transfers were greater to Eucalyptus (5-7 times) and to Casuarina (12-18 times) than in nonnodulated mycorrhizal pairs. Net transfer to Eucalyptus or to Casuarina was low in both nonnodulated nonmycorrhizal (< 0.7 mg per plant) and nonnodulated mycorrhizal pairs (< 1.1 mg per plant). In nodulated mycorrhizal pairs, net transfer to Casuarina was 26.0 mg per plant. The amount and direction of two-way mycorrhiza-mediated N transfer was increased by the presence of Pisolithus sp. and Frankia, resulting in a net N transfer from low-N-demanding Eucalyptus to high-N-demanding Casuarina.

Cyanogenic polymorphism in Eucalyptus polyanthemos Schauer subsp vestita L. Johnson and K. Hill (Myrtaceae)

Plant cyanogenesis, the release of cyanide from endogenous cyanide-containing compounds, is an effective herbivore deterrent. This paper characterises cyanogenesis in the Australian tree Eucalyptus polyanthemos Schauer subsp. vestita L. Johnson and K. Hill for the first time. The cyanogenic glucoside prunasin ((R)-mandelonitrile beta-D-glucoside) was determined to be the only cyanogenic compound in E. polyanthemos foliage. Two natural populations of E. polyanthernos showed quantitative variation in foliar prumasin concentration, varying from zero (i.e. acyanogenic) to 2.07 mg CN g(-1) dry weight in one population and from 0.17 to 1.98 mg CN g(-1) dry weight in the other. No significant difference was detected between the populations with respect to the mean prunasin concentration or the degree of variation in foliar prunasin, despite significant differences in foliar nitrogen. Variation between individuals was also observed with respect to the capacity of foliage to catabolise prunasin to form cyanide. Moreover, variation in this capacity generally correlated with the amount of prunasin in the tissue, suggesting genetic linkage between prunasin and beta-glucosidase. (C) 2002 Elsevier Science Ltd. All rights reserved.

Spore communities of arbuscular mycorrhizal fungi and mycorrhizal associations in different ecosystems, south Australia

Communities of arbuscular mycorrhizal fungi (AMF) were surveyed in different South Australian ecosystems. The soil was wet-sieved for spore extraction, followed by the determination of presence and abundance of AMF species as well as the percentage of root colonization. Mycorrhizal associations were common and there was substantial fungal diversity in different ecosystems. Spores were most abundant in the permanent pasture system and less abundant under continuous wheat. The incidence of mycorrhizal associations in different plant species and the occurrence of Arum and Paris type colonization generally conformed with previous information. Spores of seventeen AMF were verified throughout seasonal changes in 1996 and 1997 in the permanent pasture and on four host species (Lolium perenne, Plantago lanceolata, Sorghum sp. and Trifolium subterraneum) , set up with the same soils under greenhouse conditions. Glomus mosseae was the dominant spore type at all sampling times and in all trap cultures. Mycorrhizal diversity was significantly affected by different sampling times in trap cultures but not in field-collected soil. P. lanceolata, Sorghum sp. and T. subterraneum as hosts for trap cultures showed no differences in richness and diversity of AMF spores that developed in association with their roots. Abundance and diversity were lowest, however, in association with L. perenne , particularly in December 1996. Results show that the combination of spore identification from field-collected soil and trap cultures is essential to study population and diversity of AMF. The study provides baseline data for ongoing monitoring of mycorrhizal populations using conventional methods and material for the determination of the symbiotic effectiveness of AMF key members.

Effect of intercropping wheat with forage legumes on wheat production and ground cover

The use of winter legumes in southern Brazil is hindered by the slow growth of these species during establishment exposing soil surface to erosion. Introduction of these species along with spring wheat (Triticum aestivum L.) was studied as a means of increasing ground cover during their initial establishment period, without reducing wheat grain yield. Two experiments were conducted in nearby areas, one in each year. Birdsfoot trefoil (Lotus corniculatus L.), red clover (Trifolium pratense L.) cultivar Quiñequelli, white clover (T. repens L.), and arrowleaf clover (T. vesiculosum Savi) did not reduce cereal yield in either year. Wheat yield was reduced by intercropped red clover cultivar Kenland and by subclover (T. subterraneum L.) in the first year. No grain yield differences due to intercropping with any legume were detected in the second year, when rainfall was below normal. Intercropping with wheat showed to be a practical alternative to enhance ground cover at establishing forage legumes.

Determination of the impact of continuous defoliation of Lolium perenne and Trifolium repens on bacterial and fungal community structure in rhizosphere soil

To determine the effects of defoliation on microbial community structure, rhizosphere soil samples were taken pre-, and post-defoliation from the root tip and mature root regions of Trifolium repens L. and Lolium perenne L. Microbial DNA isolated from samples was used to generate polymerase chain reaction-denaturing gradient gel electrophoresis molecular profiles of bacterial and fungal communities. Bacterial plate counts were also obtained. Neither plant species nor defoliation affected the bacterial and fungal community structures in both the root tip and mature root regions, but there were significant differences in the bacterial and fungal community profiles between the two root regions for each plant. Prior to defoliation, there was no difference between plants for bacterial plate counts of soils from the root tip regions; however, counts were greater in the mature root region of L. perenne than T. repens. Bacterial plate counts for T. repens were higher in the root tip than the mature root region. After defoliation, there was no effect of plant type, position along the root or defoliation status on bacterial plate counts, although there were significant increases in bacterial plate counts with time. The results indicate that a general effect existed during maturation in the root regions of each plant, which had a greater impact on microbial community structure than either plant type or the effect of defoliation. In addition there were no generic consequences with regard to microbial populations in the rhizosphere as a response to plant defoliation.

Assessment of the effects of ozone exposure and plant competition on the reproductive ability of three therophytic clover species from Iberian pastures

Ozone (O3) phytototoxicity has been reported on a wide range of crops and wild Central European plantspecies, however no information has been provided regarding the sensitivity of plantspecies from dehesa Mediterranean therophytic grasslands in spite of their great plantspecies richness and the high O3 levels that are recorded in this area. A study was carried out in open-top chambers (OTCs) to assess the effects of O3 and competition on the reproductiveability of threecloverspecies: Trifolium cherleri, Trifolium subterraneum and Trifolium striatum. A phytometer approach was followed, therefore plants of these species were grown in mesoscosms composed of monocultures of four plants of each species, of threeplants of each species competing against a Briza maxima individual or of a single plant of each cloverspecies competing with threeB. maximaplants. Three O3 treatments were adopted: charcoal filtered air (CFA), non-filtered air (NFA) and non-filtered air supplemented with 40 nl l−1 of O3 (NFA+). The different mesocosms were exposed to the different O3 treatments for 45 days and then they remained in the open. Ozoneexposure caused reductions in the flower biomass of the threecloverspecies assessed. In the case of T. cherleri and T. subterraneum this effect was found following their exposure to the different O3 treatments during their vegetative period. An attenuation of these effects was found when the plants remained in the open. Ozone-induced detrimental effects on the seed output of T. striatum were also observed. The flower biomass of the cloverplants grown in monocultures was greater than when competing with one or threeB. maxima individuals. An increased flower biomass was found in the CFA monoculture mesocosms of T. cherleri when compared with the remaining mesocosms, once the plants were exposed in the open for 60 days. The implications of these effects on the performance of dehesa acid grasslands and for the definition of O3 critical levels is discussed