Fungal bioremediation of agricultural wastewater in a long-term treatment : biomass stabilization by immobilization strategy

Acord transformatiu CRUE-CSICFungal bioremediation emerges as an effective technology for pesticide treatment, but its successful implementation depends on overcoming the problem of microbial contamination. In this regard, fungal immobilization on wood seems to be a promising strategy, but there are two main drawbacks: the predominant removal of pesticides by sorption and fungal detachment. In this study, agricultural wastewater with pesticides was treated by Trametes versicolor immobilized on wood chips in a rotary drum bioreactor (RDB) for 225 days, achieving fungal consolidation and high pesticide biodegradation through two main improvements: the use of a more favorable substrate and the modification of operating conditions. Fungal community dynamic was assessed by denaturing gradient gel electrophoresis (DGGE) analysis and subsequent prominent band sequencing, showing a quite stable community in the RDB, mainly attributed to the presence of T. versicolor. Pesticide removals were up to 54 % diuron and 48 % bentazon throughout the treatment. Afterwards, pesticide-contaminated wood chips were treated by T. versicolor in a solid biopile-like system. Hence, these results demonstrate that the microbial contamination constraint has definitely been overcome, and fungal bioremediation technology is ready to be implemented on a larger scale

Stratification strength and light climate explain variation in chlorophyll a at the continental scale in a European multilake survey in a heatwave summer.

To determine the drivers of phytoplankton biomass, we collected standardized morphometric, physical, and biological data in 230 lakes across the Mediterranean, Continental, and Boreal climatic zones of the European continent. Multilinear regression models tested on this snapshot of mostly eutrophic lakes (median total phosphorus [TP] = 0.06 and total nitrogen [TN] = 0.7 mg L 1), and its subsets (2 depth types and 3 climatic zones), show that light climate and stratification strength were the most significant explanatory variables for chlorophyll a (Chl a) variance. TN was a significant predictor for phytoplankton biomass for shallow and continental lakes, while TP never appeared as an explanatory variable, suggesting that under high TP, light, which partially controls stratification strength, becomes limiting for phytoplankton development. Mediterranean lakes were the warmest yet most weakly stratified and had significantly less Chl a than Boreal lakes, where the temperature anomaly from the long-term average, during a summer heatwave was the highest (+4 C) and showed a significant, exponential relationship with stratification strength. This European survey represents a summer snapshot of phytoplankton biomass and its drivers, and lends support that light and stratification metrics, which are both affected by climate change, are better predictors for phytoplankton biomass in nutrient-rich lakes than nutrient concentrations and surface temperature

Complete sequence and genomic annotation of carrot torradovirus 1

Carrot torradovirus 1 (CaTV1) is a new member of the genus Torradovirus within the family Secoviridae. CaTV1 genome sequences were obtained from a previous next-generation sequencing (NGS) study and were compared to other members and tentative new members of the genus. The virus has a bipartite genome, and RACE was used to amplify and sequence each end of RNA1 and RNA2. As a result, RNA1 and RNA2 are estimated to contain 6944 and 4995 nucleotides, respectively, with RNA1 encoding the proteins involved in virus replication, and RNA2 encoding the encapsidation and movement proteins. Sequence comparisons showed that CaTV1 clustered within the non-tomato-infecting torradoviruses and is most similar to motherwort yellow mottle virus (MYMoV). The nucleotide sequence identities of the Pro-Pol and coat protein regions were below the criteria established by the ICTV for demarcating species, confirming that CaTV1 should be classified as a member of a new species within the genus Torradovirus

A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants

The production of plant helical virus-like particles (VLPs) via plant-based expression has been problematic with previous studies suggesting that an RNA scaffold may be necessary for their efficient production. To examine this, we compared the accumulation of VLPs from two potexviruses, papaya mosaic virus and alternanthera mosaic virus (AltMV), when the coat proteins were expressed from a replicating potato virus X- based vector (pEff) and a non-replicating vector (pEAQ-HT). Significantly greater quantities of VLPs could be purified when pEff was used. The pEff system was also very efficient at producing VLPs of helical viruses from different virus families. Examination of the RNA content of AltMV and tobacco mosaic virus VLPs produced from pEff revealed the presence of vector-derived RNA sequences, suggesting that the replicating RNA acts as a scaffold for VLP assembly. Cryo-EM analysis of the AltMV VLPs showed they had a structure very similar to that of authentic potexvirus particles. Thus, we conclude that vectors generating replicating forms of RNA, such as pEff, are very efficient for producing helical VLPs

Influence of phenological barriers and habitat differentiation on the population genetic structure of the balearic endemic Rhamnus ludovici-salvatoris Chodat and R. alaternus L

[EN] Rhamnus ludovici-salvatoris, endemic to the Gymnesian Islands, coexists with the related and widespread R. alaternus in Mallorca and Menorca. In both species, the population genetic structure using RAPD, and flowering during a 3-year period to check for possible phenological barriers, were analyzed. Rhamnus ludovici-salvatoris showed lower genetic diversity and stronger population structure than R. alaternus, the Cabrera population being less diverse and the most differentiated. Rhamnus ludovici-salvatoris flowered one month later, although flowering of both species coincided sporadically. These congeners seem to have diverged through isolation by time and differentiation in habitat. The population genetic structure of R. ludovici-salvatoris could mainly be due to the existence of small populations on the one hand, and a gene flow caused by rare hybridization events on the other, which may also explain the presence of morphologically intermediate individuals in Menorca. The conservation of R. ludovici-salvatoris populations may include population reinforcements and other in situ interventions.Ferriol Molina, M.; Llorens García, L.; Gil, L.; Boira Tortajada, H. (2009). Influence of phenological barriers and habitat differentiation on the population genetic structure of the balearic endemic Rhamnus ludovici-salvatoris Chodat and R. alaternus L. Plant Systematics and Evolution. 277(1-2):105-116. doi:10.1007/s00606-008-0110-3S1051162771-2Affre L, Thompson JD, Debussche M (1997) Genetic structure of continental and island populations of the Mediterranean endemic Cyclamen balearicum (Primulaceae). Amer J Bot 84(4): 437–451BOIB (2005) Decreto 75/2005. BOIB 106: 29–32Bolmgren K, Oxelman B (2004) Generic limits in Rhamnus L. s.l. (Rhamnaceae) inferred from nuclear and chloroplast DNA sequence phylogenies. Taxon 53(2):383–390Bolòs O, Molinier R (1958) Recherches phytosociologiques dans l’île de Majorque. Collectanea Botanica 34:699–865Cardona MA (1979) Consideracions sobre l’endemisme i l’origen de la flora de las Illes Balears. Butlletí del Institut Catalá de Historia Natural 44 (Sec. Bot. 3):7–15Cardona MA, Contandriopoulos J (1979) Endemism and evolution in the islands of the Western Mediterranean. In: Bramwell D (ed) Plants and islands. Academic Press, London, pp 133–169Chodat L (1924) Contributions à la Géo-Botanique de Majorque. PhD Thesis, Université de Genève—Institut de Botanique, SwitzerlandCollins D, Mill RR, Moller M (2003) Species separation of Taxus baccata, T. canadensis, and T. cuspidata (Taxaceae) and origins of their reputed hybrids inferred from RAPD and cpDNA data. Amer J Bot 90(2):175–182Cronk QCB (1997) Islands: stability, diversity, conservation. Biodivers Conserv 6(3):477–493Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Ducarme V, Wesselingh RA (2005) Detecting hybridization in mixed populations of Rhinanthus minor and Rhinanthus angustifolius. Folia Geobot 40(2/3):151–161Englishloeb GM, Karban R (1992) Consequences of variation in flowering phenology for seed head herbivory and reproductive success in Erigeron glaucus (Compositae). Oecologia 89:588–595Gautier F, Caluzon G, Suk JP, Violanti D (1994) Age et durée de la crise de salinité Messinienne. Comptes Rendus de l’Académie des Sciences de Paris 318:1103–1109Gerard PR, Fernandez-Manjarres JF, Frascaria-Lacoste N (2006) Temporal cline in a hybrid zone population between Fraxinus excelsior L. and Fraxinus angustifolia Vahl. Molec Ecol 15:3655–3667Gil L, Llorens L, Tébar FJ, Costa M (1995) La vegetación de la isla de Cabrera. In: Guía de la excursión geobotánica de las XV Jornadas de Fitosociología. Datos sobre la vegetación de Cabrera. Palma de Mallorca: Universitat de les Illes Balears, pp 51–77Gulías J, Flexas J, Abadía A, Medrano H (2002) Photosynthetic responses to water deficit in six Mediterranean sclerophyll species: possible factors explaining the declining distribution of Rhamnus ludovici-salvatoris, and endemic Balearic species. Tree Physiol 22:687–697Gulías J, Traveset A, Riera N, Mus M (2004) Critical stages in the recruitment process of Rhamnus alaternus L. Ann Bot 93:723–731Gustafsson S, Sjögren-Gulve P (2002) Genetic diversity in the rare orchid, Gymnadenia odoratissima and a comparison with the more common congener, G. conopsea. Conserv Genet 3:225–234Gustafsson S (2003) Population genetic analyses in the orchid genus Gymnadenia—a conservation genetic perspective. PhD Thesis, Uppsala University, SwedenGustafsson S, Lönn M (2003) Genetic differentiation and habitat preference of flowering-time variants within Gymnadenia conopsea. Heredity 91:284–292Harris W (1996) Genecological aspects of flowering patterns of populations of Kunzea ericoides and K. sinclairii (Myrtaceae). New Zealand J Bot 34:333–354Hendry AP, Dray T (2005) Population structure attributable to reproductive time: isolation by time and adaptation by time. Molec Ecol 14:901–916Hosokawa K, Minami M, Kawahara K, Nakamura I, Shibata T (2000) Discrimination among three species of medicinal Scutellaria plants using RAPD markers. Pl Med 66:270–272Huang Z, Liu L, Zhou T, Ju B (2005) Effects of environmental factors on the population genetic structure in chukar partridge (Alectoris chukar). J Arid Environ 62:427–434Juan A, Crespo MB, Cowan RS, Lexer C, Fay F (2004) Patterns of variability and gene flow in Medicago citrina, an endangered endemic of islands in the western Mediterranean, as revealed by amplified fragment length polymorphism (AFLP). Molec Ecol 13:2679–2690Krijgsman W, Hilgen FJ, Raffi I, Sierro FJ, Wilson DS (1999) Chronology, causes and progression of the Messinian salinity crisis. Nature 400:652–655Lamont BB, He T, Enright NJ, Krauss SL, Miller BP (2003) Anthropogenic disturbance promotes hybridization between Banksia species by altering their biology. J Evol Biol 16:551–557Lennartsson T (1997) Seasonal differentiation—a conservative reproductive barrier in two grassland Gentianella (Gentianaceae) species. Pl Syst Evol 208:45–69Martinez-Solis I, Iranzo J, Estrelles E, Ibars AM (1993) Leaf domatia in the section Alaternus (Miller) DC. of the genus Rhamnus (Rhamnaceae). Bot J Linn Soc 112:311–318McIntosh ME (2002) Flowering phenology and reproductive output in two sister species of Ferocactus (Cactaceae). Pl Ecol 159:1–13Nei M (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA 70:3321–3323Nei M (1978) Estimation of average heterozigosity and genetic distance from a small number of individuals. Genetics 89:583–590Nei M, Li W (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 79:5269–5273Nybom H, Bartish IV (2000) Effects of life history traits and sampling strategies on genetic diversity estimates obtained with RAPD markers in plants. Perspect Pl Ecol Evol Syst 3(2):93–114Oostermeijer JGB, Luijten SH, Ellis-Adam AC, den Nijs JCM (2002) Future prospects for the rare, late-flowering Gentianella germanica and Gentianopsis ciliata in Dutch nutrient-poor calcareous grasslands. Biol Conserv 104:339–350Pease CM, Lande R, Bull JJ (1989) A model of population growth, dispersal and evolution in a changing environment. Ecology 70(6):1657–1664Perron M, Gordon AG, Bousquet J (1995) Species-specific RAPD fingerprints for the closely related Picea mariana and P. rubens. Theor Appl Genet 91:142–149Pierce S, Ceriani RM, Villa M, Cerabolini B (2006) Quantifying relative extinction risks and targeting intervention for the orchid flora of a natural park in the European prealps. Conserv Biol 20(6):1804–1810Richardson JE, Fay MF, Cronk QCB, Bowman D, Chase MW (2000) A phylogenetic analysis of Rhamnaceae using rbcL and trnL-F plastid DNA sequences. Amer J Bot 87(9):1309–1324Roselló JA, Sáez L (2000) Index Balearicum: an annotated check-list of the vascular plants described from the Balearic Islands. Collect Bot 25(1):3–203Roselló JA, Cebrián MC, Mayol M (2002) Testing taxonomic and biogeographical relationships in a narrow mediterranean endemic complex (Hippocrepis balearica) using RAPD markers. Ann Bot 89:321–327Sales E, Nebauer SG, Mus M, Segura J (2001) Population genetic study in the Balearic plant species Digitalis minor (Scrophulariaceae) using RAPD markers. Amer J Bot 88(10):1750–1759Sherwin WB, Moritz C (2000) Managing and monitoring genetic erosion. In: Young AG, Clarke GM (eds) Genetics, demography and viability of fragmented populations. Cambridge University Press, Cambridge, pp 9–34Sneath PHA, Sokal RR (1973) Numerical taxonomy. Freeman and Co., San FranciscoTraveset A, Gulías J, Riera N, Mus M (2003) Transition probabilities from pollination to establishment in a rare dioecious shrub species (Rhamnus ludovici-salvatoris) in two habitats. J Ecol 91:427–437Tutin TG, Heywood VH, Burges NA, Valentine DH, Walters SM, Webb DA (eds) (2001) Flora Europaea, vol 2. Rosaceae to Umbelliferae. Cambridge University Press, CambridgeWright S (1931) Evolution in Mendelian populations. Genetics 16:97–159Zimmerman M (1980a) Reproduction in Polemonium: pre-dispersal seed predation. Ecology 61:502–506Zimmerman M (1980b) Reproduction in Polemonium: competition for pollinators. Ecology 61:497–50

Environmental-dependent proline accumulation in plants living on gypsum soils

[EN] Biosynthesis of proline¿or other compatible solutes¿is a conserved response of all organisms to different abiotic stress conditions leading to cellular dehydration. However, the biological relevance of this reaction for plant stress tolerance mechanisms remains largely unknown, since there are very few available data on proline levels in stress-tolerant plants under natural conditions. The aim of this work was to establish the relationship between proline levels and different environmental stress factors in plants living on gypsum soils. During the 2-year study (2009¿2010), soil parameters and climatic data were monitored, and proline contents were determined, in six successive samplings, in ten taxa present in selected experimental plots, three in a gypsum area and one in a semiarid zone, both located in the province of Valencia, in south-east Spain. Mean proline values varied significantly between species; however, seasonal variations within species were in many cases even wider, with the most extreme differences registered in Helianthemum syriacum (almost 30 lmol g-1 of DW in summer 2009, as compared to ca. 0.5 in spring, in one of the plots of the gypsum zone). Higher proline contents in plants were generally observed under lower soil humidity conditions, especially in the 2009 summer sampling preceded by a severe drought period. Our results clearly show a positive correlation between the degree of environmental stress and the proline level in most of the taxa included in this study, supporting a functional role of proline in stress tolerance mechanisms of plants adapted to gypsum. However, the main trigger of proline biosynthesis in this type of habitat, as in arid or semiarid zones, is water deficit, while the component of ¿salt stress¿ due to the presence of gypsum in the soil only plays a secondary role.This work has been supported by the Spanish Ministry of Science and Innovation (Project CGL2008-00438/BOS), with contribution from the European Regional Development Fund.Boscaiu, M.; Bautista Carrascosa, I.; Lidón Cerezuela, AL.; Llinares Palacios, JV.; Lull, C.; Donat-Torres, M.; Mayoral García-Berlanga, O.... (2013). Environmental-dependent proline accumulation in plants living on gypsum soils. Acta Physiologiae Plantarum. 35:2193-2204. https://doi.org/10.1007/s11738-013-1256-3S2193220435Alvarado JJ, Ruiz JM, López-Cantarero I, Molero J, Romero L (2000) Nitrogen metabolism in five plant species characteristic of gypsiferous soils. J Plant Physiol 156:612–616Ashraf M, Foolad MR (2007) Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot 59:206–216Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207Briens M, Larher F (1982) Osmoregulation in halophytic higher plants: a comparative study of soluble carbohydrates, polyols, betaines and free proline. Plant, Cell Environ 5:287–292Burriel F, Hernando V (1947) Nuevo método para determinar el fósforo asimilable en los suelos. Anales de Edafología y Fisiología Vegetal 9:611–622Caballero I, Olano JM, Loidi J, Escudero A (2003) Seed bank structure along a semi-arid gypsum gradient in Central Spain. J Arid Environ 55:287–299Escudero A, Carnes LF, Pérez García F (1997) Seed germination of gypsophytes and gypsovags in semi-arid central Spain. J Arid Environ 36:487–497Escudero A, Somolinos RC, Olano JM, Rubio A (1999) Factors controlling the establishment of Helianthemum squamatum, an endemic gypsophite of semi-arid Spain. J Ecol 87:290–302FAO (1990) Management of gypsiferous soils. FAO Soils Bull 62Ferriol M, Pérez I, Merle H, Boira H (2006) Ecological germination requirements of the aggregate species Teucrium pumilum (Labiatae) endemic to Spain. Plant Soil 284:205–216Flowers TJ, Colmer TD (2008) Salinity tolerance in halophytes. New Phytol 179:945–963Flowers TJ, Troke PF, Yeo AR (1977) The mechanism of salt tolerance in halophytes. Ann Rev Plant Physiol 28:89–121Gil R, Lull C, Boscaiu M, Bautista I, Lidón A, Vicente O (2011) Soluble carbohydrates as osmolytes in several halophytes from a Mediterranean salt marsh. Not Bot Horti Agrobo 39(2):9–17Grigore MN, Boscaiu M, Vicente O (2011) Assessment of the relevance of osmolyte biosynthesis for salt tolerance of halophytes under natural conditions. Eur J Plant Sci Biotech 5:12–19Hare PD, Cress WA, Van Standen J (1998) Dissecting the roles of osmolyte accumulation during stress. Plant Cell Environ 21:535–553Keeney DR, Nelson DW (1982) Nitrogen inorganic forms. In: Page AL et al (eds) Methods of soil analysis, part 2: chemical and microbiological properties. Soil Science Society of America, Madison, pp 643–698Knudsen D, Peterson GA, Pratt PF (1982) Lithium, Sodium and Potassium. In: Page AL et al (eds) Methods of soil analysis, part 2: chemical and microbiological properties. Soil Science Society of America, Madison, pp 225–246Kuo S (1996) Phosphorus. In: Spark DL (ed) Methods of soil analysis: chemical methods, part 3. Soil Science Society of America, Madison, pp 869–919Martens H, Maes T (1989) Multivariate calibration. Wiley, New York, pp 97–108Martínez-Duro E, Ferrandis P, Escudero A, Luzuriaga AL, Herranz JM (2010) Secondary old-field succession in an ecosystem with restrictive soils: does time from abandonment matter? Appl Veg Sci 13:234–248Meyer SE (1986) The ecology of gypsophile endemism in the eastern Mojave desert. Ecology 67:1303–1313Meyer SE, García-Moya E (1989) Plant community patterns and soil moisture regime in gypsum grasslands of north central Mexico. J Arid Environ 16:147–155Meyer SE, García-Moya E, Lagunes-Espinoza LC (1992) Topographic and soil surface effects on gypsophile plant community patterns in central Mexico. J Veg Sci 3:429–438Moruno F, Soriano P, Vicente O, Boscaiu M, Estrelles E (2011) Opportunistic germination behaviour of Gypsophila (Caryophyllaceae) in two priority habitats from semi-arid Mediterranean steppes. Not Bot Horti Agrobo 39(1):18–23Mota JF, Sánchez Gómez P, Merlo Calvente ME, Catalán Rodríguez P, Laguna Lumbreras E, de la Cruz Rot M, Navarro Reyes FB, Marchal Gallardo F, Bartolomé Esteban C, Martínez Labarga JM, Sainz Ollero H, Valle Tendero F, Serra Laliga L, Martínez Hernández F, Garrido Becerra JA, Pérez García FJ (2009) Aproximación a la checklist de los gipsófitos ibéricos. Anales de Biología 31:71–80Murakeözy ÉP, Nagy Z, Duhazé C, Bouchereau A, Tuba Z (2003) Seasonal changes in the levels of compatible osmolytes in three halophytic species of inland saline vegetation in Hungary. J Plant Physiol 160:395–401Nelson DW, Sommers LE (1982) Total carbon, organic carbon, and organic matter. In: Page AL et al (eds) Methods of soil analysis, part 2: chemical and microbiological properties. Soil Science Society of America, Madison, pp 539–577Palacio S, Escudero A, Montserrat-Martí G, Maestro M, Milla R, Albert M (2007) Plants living on gypsum: beyond the specialist model. Ann Bot 99:333–343Parsons RF (1977) Gypsophily in plants—a review. Am Midl Nat 96:1–20Pueyo Y, Alados CL, Maestro M, Komac B (2007) Gypsophile vegetation patterns under a range of soil properties induced by topographical position. Plant Ecol 189:301–311Rivas-Martínez S, Rivas-Sáenz S (2009) Worldwide Bioclimatic Classification System. Phytosociological Research Center, Complutense University of Madrid, Spain. http://www.globalbioclimatics.org/ . Accessed 15 Nov 2012Romão RL, Escudero A (2005) Gypsum physical soil crusts and the existence of gypsophytes in semi-arid central Spain. Plant Ecol 181:127–137Rubio A, Escudero A (2000) Small-scale spatial soil-plant relationship in semi-arid gypsum environment. Plant Soil 220:139–150Ruíz JM, López-Cantarero I, Rivero RM, Romero L (2003) Sulphur phytoaccumulation in plant species characteristic of gypsiferous soils. Int J Phytorem 5:203–210Szabados L, Savouré A (2010) Proline: a multifunctional amino acid. Trends Plant Sci 15:89–97Szabados L, Kovács H, Zilberstein A, Bouchereau A (2011) Plants in extreme environments: importance of protective compounds in stress tolerance. Adv Bot Res 57:105–150Tecator Application Note (1984) AN 5226: Determination of ammonium in 2 M KCl soil extracts by FIAstar 5000. AN 5201: Determination of the sum of nitrate and nitrite in water by FIAstar 5000. (Adapted for 2 M KCl soil extracts)Tipirdamaz R, Gagneul D, Duhazé C, Aïnouche A, Monnier C, Özkum D, Larher F (2006) Clustering of halophytes from an inland salt marsh in Turkey according to their ability to accumulate sodium and nitrogenous osmolytes. Environ Exp Bot 57:139–153Verheye WH, Boyadgiev TG (1997) Evaluating the land use potential of gypsiferous soils from field pedogenic characteristics. Soil Use Manage 13:97–103Vicente O, Boscaiu M, Naranjo MA, Estrelles E, Bellés JM, Soriano P (2004) Responses to salt stress in the halophyte Plantago crassifolia (Plantaginaceae). J Arid Environ 58:463–481Yancey PH (2005) Organic osmolytes as compatible, metabolic and counteracting cytoprotectants in high osmolarity and other stresses. J Exp Biol 208:2819–283