Comparative epigenomics in distantly related teleost species identifies conserved cis-regulatory nodes active during the vertebrate phylotypic period

The complex relationship between ontogeny and phylogeny has been the subject of attention and controversy since von Baer’s formulations in the 19th century. The classic concept that embryogenesis progresses from clade general features to species-specific characters has often been revisited. It has become accepted that embryos from a clade show maximum morphological similarity at the so-called phylotypic period (i.e., during mid-embryogenesis). According to the hourglass model, body plan conservation would depend on constrained molecular mechanisms operating at this period. More recently, comparative transcriptomic analyses have provided conclusive evidence that such molecular constraints exist. Examining cis-regulatory architecture during the phylotypic period is essential to understand the evolutionary source of body plan stability. Here we compare transcriptomes and key epigenetic marks (H3K4me3 and H3K27ac) from medaka (Oryzias latipes) and zebrafish (Danio rerio), two distantly related teleosts separated by an evolutionary distance of 115-200 Myr. We show that comparison of transcriptome profiles correlates with anatomical similarities and heterochronies observed at the phylotypic stage. Through comparative epigenomics, we uncover a pool of conserved regulatory regions (approximate to 700), which are active during the vertebrate phylotypic period in both species. Moreover, we show that their neighboring genes encode mainly transcription factors with fundamental roles in tissue specification. We postulate that these regulatory regions, active in both teleost genomes, represent key constrained nodes of the gene networks that sustain the vertebrate body plan

Deployment of mating disruption dispensers before and after first seasonal male flights for the control of Aonidiella aurantii in citrus

The rejection of citrus fruit caused by infestations of the California red scale (CRS), Aonidiella aurantii (Maskell) (Hemiptera: Diaspididae), raises concerns about its management. This fact has led to the introduction of new integrated control methods in citrus orchards, including the implementation of techniques based on pheromones. Previous works described efficient mating disruption pheromone dispensers to control A. aurantii in the Mediterranean region. The main aims of the present study were to adjust the timing of dispenser applications and study the importance of controlling the early first generation of A. aurantii by testing two different application dates: before and after the first CRS male flight. The efficacy of the different mating disruption strategies was tested during 2010 in an experimental orchard and these results were confirmed during 2011 in a commercial citrus farm. Results showed that every mating disruption strategy achieved significantly lower male captures in monitoring pheromone traps compared with untreated plots, as well as mean fruit infestation reductions of about 80 %. The control of the first CRS generation is not essential for achieving a good efficacy as demonstrated in two locations with different pest pressure. The late application of MD dispensers before the second CRS male flight has proven to be effective, suggesting a new advantageous way to apply mating disruption.The authors want to thank Fernando Alfaro from Denia, Antonio Caballero, and Javier Macias from Rio Tinto Fruit S.A. (Huelva, Spain) for field support. We also thank Ecologia y Proteccion Agricola SL for the pheromone supply. This work has been funded by the Spanish Ministry of Science and Innovation (project AGL2009-10725) and Agroalimed Foundation. The translation of this paper was funded by the Universidad Politecnica de Valencia (Spain).Vacas González, S.; Alfaro Cañamás, C.; Primo Millo, J.; Navarro-Llopis, V. (2015). Deployment of mating disruption dispensers before and after first seasonal male flights for the control of Aonidiella aurantii in citrus. Journal of Pest Science. 88(2):321-329. https://doi.org/10.1007/s10340-014-0623-1S321329882Avidov Z, Balshin M, Gerson U (1970) Studies on Aphytis coheni, a parasite of the California red scale, Aonidiella aurantii in Israel. Biocontrol 15:191–207Barzakay I, Hefetz A, Sternlicht M, Peleg BA, Gokkes M, Singer G, Geffen D, Kronenberg S (1986) Further field trials on management of the California red scale, Aonidiella aurantii, by mating disruption with its sex-pheromone. Phytoparasitica 14:160–161Bedford ECG (1996) Problems which we face in bringing red scale, Aonidiella aurantii (Maskell), under biological control in citrus in South Africa. Proc Int Soc Citriculture 1:485–492Campos-Rivela JM, Martínez-Ferrer MT, Fibla-Queralt JM (2012) Population dynamics and seasonal trend of California red scale (Aonidiella aurantii Maskell) in citrus in Northern Spain. Span J Agric Res 10:198–208Collins PJ, Lambkin TM, Bodnaruk P (1994) Suspected resistance to methidation in Aonidiella aurantii (Maskell) (Homoptera: diaspididae) from Queensland. J Aust Entomol Soc 33:325–326Corma A, Muñoz-Pallares J, Primo-Yufera E (1999) Production of semiochemical emitters having a controlled emission speed which are based on inorganic molecular sieves. World Patent WO9944420Corma A, Muñoz-Pallares J, Primo-Yufera E (2000) Emitter of semiochemical substances supported on a sepiolite, preparation process and applications. World Patent WO0002448DeBach P (1959) New species and strains of Aphytis (Hymenoptera: Eulophidae) parasitic on the California red scale, Aonidiella aurantii (Mask.), in the Orient. Ann Entomol Soc Am 52:354–362DeBach P, Argyriou L (1967) The colonization and success in Greece of some imported Aphytis spp. (Hymenoptera: Aphelinidae) parasitic on citrus scale insects (Homoptera: Diaspididae). Biocontrol 12:325–342Desneux N, Decourtye A, Delpuech JM (2007) The sublethal effects of pesticides on beneficial arthropods. Ann Rev Entomol 52:81–106Diari Oficial de la Comunitat Valenciana (DOCV) (2008) DOCV no. 5901, 26. Resolution 27 October 2008 of Consellería de Agricultura, Pesca y Alimentación; November 2008. http://www.docv.gva.es/datos/2008/11/26/pdf/2008_13692.pdfDomínguez-Ruiz J, Sanchis J, Navarro-Llopis V, Primo J (2008) A new long-life trimedlure dispenser for Mediterranean fruit fly. J Econ Entomol 101:1325–1330Eliahu M, Blumberg D, Horowitz AR, Ishaaya I (2007) Effect of pyriproxyfen on developing stages and embryogenesis of California red scale (CRS), Aonidiella aurantii. Pest Manag Sci 63:743–746Furness G, Buchanan G, George R, Richardson N (1983) A history of the biological and integrated control of red scale, Aonidiella aurantii on citrus in the lower Murray Valley of Australia. Biocontrol 28:99–212Grafton-Cardwell EE, Gu P (2003) Conserving vedalia beetle, Rodolia cardinalis (Mulsant) (Coleoptera : Coccinellidae), in citrus: a continuing challenge as new insecticides gain registration. J Econ Entomol 96:1388–1398Grafton-Cardwell EE, Reagan CA (1995) Selective use of insecticides for control of armored scale (Homoptera: Diaspididae) in San-Joaquin Valley California citrus. J Econ Entomol 88:1717–1725Grafton-Cardwell EE, Vehrs SLC (1995) Monitoring for organophosphate-resistant and carbamate-resistant armored scale (Homoptera: Diaspididae) in San-Joaquin Valley citrus. J Econ Entomol 88:495–504Grafton-Cardwell EE, Lee JE, Stewart JR, Olsen KD (2006) Role of two insect growth regulators in integrated pest management of citrus scales. J Econ Entomol 99:733–744Grout TG, Richards GI (1991a) Effect of buprofezin applications at different phenological times on California red scale (Homoptera: Diaspididae). J Econ Entomol 84:1802–1805Grout TG, Richards GI (1991b) Value of pheromone traps for predicting infestations of red scale, Aonidiella aurantii (Maskell) (Homoptera: Diaspididae), limited by natural enemy activity and insecticides used to control citrus thrips, Scirtothrips aurantii Faure (Thysanoptera: Thripidae). J Appl Entomol 111:20–27Grout TG, Du Toit WJ, Hofmeyr JH, Richards GI (1989) California red scale (Homoptera: Diaspididae) phenology on citrus in South Africa. J Econ Entomol 82:793–798Hefetz A, Kronengerg S, Peleg BA, Bar-zakay I (1988) Mating disruption of the California red scale Aonidiella aurantii (Homoptera: Diaspididae). In: Proceeding 6th International Citrus Congress, Tel Aviv (Israel), pp 1121–1127Hernández-Penadés P, Rodríguez-Reina JM, García-Marí F (2002) Umbrales de tratamiento para cóccidos diaspídidos en cítricos. Bol San Veg Plagas 28:469–478Hothorn T, Bretz F, Westfall P (2008) Simultaneous Inference in General Parametric Models. Biometrical J 50:346–363Ioratti C, Anfora G, Tasin M, De Cristofaro A, Witzgall P, Lucchi A (2011) Chemical ecology and management of Lobesia botrana (Lepidoptera: Tortricidae). J Econ Entomol 104:1125–1137Kehat M, Anshelevich L, Harel M, Dunkelblum E (1995) Control of the codling moth (Cydia pomonella) in apple and pear orchards in Israel by mating disruption. Phytoparasitica 23:285–296Kennett CE, Hoffmann RW (1985) Seasonal development of the California red scale (Homoptera: Diaspididae) in San Joaquin Valley citrus based on degree-day accumulation. J Econ Entomol 78:73–79Levitin E, Cohen E (1998) The involvement of acetylcholinesterase in resistance of the California red scale shape Aonidiella aurantii to organophosphorus pesticides. Entomol Exp Appl 88:115–121Lykouressis D, Perdikis D, Samartzis D, Fantinou A, Toutouzas S (2005) Management of the pink bollworm Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae) by mating disruption in cotton fields. Crop Prot 24:177–183McLaren IW, Buchanan GA (1973) Parasitism by Aphytis chrysomphali Mercet and A. melinus Debach of Californian red scale, Aonidiella aurantii (Maskell), in relation to seasonal availability of suitable stages of the scale. Austr J Zool 21:111–117Moreno DS, Kennett CE (1985) Predictive year-end California red scale (Homoptera: Diaspididae) orange fruit infestations based on catches of males in the San-Joaquin Valley. J Econ Entomol 78:1–9Moreno DS, Luck RF (1992) Augmentative releases of Aphytis melinus (Hymenoptera: Aphelinidae) to suppress California red scale (Homoptera: Diaspididae) in southern California lemon orchards. J Econ Entomol 85:1112–1119Pekas A, Aguilar A, Tena A, García-Marí F (2010) Influence of host size on parasitism by Aphytis chrysomphali and A. melinus (Hymenoptera: Aphelinidae) in Mediterranean populations of California red scale Aonidiella aurantii (Hemiptera: Diaspididae). Biol Control 55:132–140Rill S, Grafton-Cardwell EE, Morse JG (2007) Effects of pyriproxyfen on California red scale (Hemiptera: Diaspididae) development and reproduction. J Econ Entomol 100:1435–1443Rodrigo E, Troncho P, García-Marí F (1996) Parasitoids (Hymenoptera: Aphelinidae) of three scale insects (Homoptera: Diaspididae) in a citrus grove in Valencia, Spain. Entomophaga 41:77–94Roelofs WL, Gieselmann MJ, Cardé AM, Tashiro H, Moreno DS, Henrick CA, Anderson RJ (1977) Sex-pheromone of California red scale, Aonidiella aurantii. Nature 26:698–699Rongai D, Cerato C, Lazzeri L, Palmieri S, Patalano G (2008) Vegetable oil formulation as biopesticide to control California red scale (Aonidiella aurantii Maskell). J Pest Sci 81:179–185Sorribas JJ, Rodríguez R, Rodrigo E, García-Marí F (2008) Niveles de parasitismo y especies de parasitoides del piojo rojo de california Aonidiella aurantii (Hemiptera: Diaspididae) en cítricos de la Comunidad Valenciana. Bol San Veg Plagas 34:201–210Sorribas J, van Baaren J, Garcia-Marí F (2012) Effects of climate on the introduction, distribution and biotic potential of parasitoids: applications to biological control of California red scale. Biol Control 62:103–112Staten RT, Flint HM, Weddle RC, Quintero E, Zarate RE, Finell CM, Hernandes M, Yamamoto A (1987) Pink bollworm (Lepidoptera: Gelechiidae): large-scale field trials with a high-rate gossyplure formulation. J Econ Entomol 80:1267–1271Tashiro H, Chambers DL (1967) Reproduction in the California Red Scale, Aonidiella aurantii (Homoptera: Diaspididae). I. Discovery and extraction of a female sex pheromone. Ann Entomol Soc Am 60:1166–1170Tena A, Llácer E, Urbaneja A (2013) Biological control of a non-honeydew producer mediated by a distinct hierarchy of honeydew quality. Biol Control 67:117–122University of California (1991) Integrated pest management for citrus. University of California, BerkeleyVacas S, Alfaro C, Navarro-Llopis V, Primo J (2009) The first account of the mating disruption technique for the control of California red scale Aonidiella aurantii Maskell (Homoptera: Diaspididae) using new biodegradable dispensers. Bull Entomol Res 99:415–423Vacas S, Alfaro C, Navarro-Llopis V, Primo J (2010) Mating disruption of California red scale, Aonidiella aurantii Maskell (Homoptera: Diaspididae), using biodegradable mesoporous pheromone dispensers. Pest Manag Sci 66:745–751Vacas S, Vanaclocha P, Alfaro C, Primo J, Verdú MJ, Urbaneja A, Navarro-Llopis V (2011) Mating disruption for the control of Aonidiella aurantii Maskell (Hemiptera: Diaspididae) may contribute to increased effectiveness of natural enemies. Pest Manag Sci 68:142–148Vanaclocha P, Vacas S, Alfaro C, Primo J, Verdú MJ, Navarro-Llopis V, Urbaneja A (2012) Life history parameters and scale-cover surface area of Aonidiella aurantii are altered in a mating disruption environment: implications for biological control. Pest Manag Sci 68:1092–1097Vanaclocha P, Vidal-Quist C, Oheix S, Montón H, Planes L, Catalán J, Tena A, Verdú MJ, Urbaneja A (2013) Acute toxicity in laboratory tests of fresh and aged residues of pesticides used in citrus on the parasitoid Aphytis melinus. J Pest Sci 86:329–336Yarom I, Blumberg D, Ishaaya I (1988) Effects of buprofezin on California red scale (Homoptera: Diaspididae) and Mediterranean black scale (Homoptera: Coccidae). J Econ Entomol 81:1581–1585Yust HR, Nelson HD, Busbey RL (1943) Comparative susceptibility of two strains of California red scale to HCN, with special reference to the inheritance of resistance. J Econ Entomol 36:744–74

Xenopus as a Model System for the Study of GOLPH2/GP73 Function: Xenopus golph2 Is Required for Pronephros Development

GOLPH2 is a highly conserved protein. It is upregulated in a number of tumors and is being considered as an emerging biomarker for related diseases. However, the function of GOLPH2 remains unknown. The Xenopus model is used to study the function of human proteins. We describe the isolation and characterization of Xenopus golph2, which dimerizes and localizes to the Golgi in a manner similar to human GOLPH2. Xenopus golph2 is expressed in the pronephros during early development. The morpholino-mediated knockdown of golph2 results in edema formation. Additionally, Nephrin expression is enhanced in the glomus, and the expression of pronephric marker genes, such as atp1b1, ClC-K, NKCC2, and NBC1, is diminished in the tubules and duct. Expression patterns of the transcription factors WT1, Pax2, Pax8, Lim1, GATA3, and HNF1β are also examined in the golph2 knockdown embryos, the expression of WT1 is increased in the glomus and expanded laterally in the pronephric region. We conclude that the deletion of golph2 causes an increase in the expression of WT1, which may promote glomus formation and inhibit pronephric tubule differentiation

Generation of Immortal Cell Lines from the Adult Pituitary: Role of cAMP on Differentiation of SOX2-Expressing Progenitor Cells to Mature Gonadotropes

The pituitary is a complex endocrine tissue composed of a number of unique cell types distinguished by the expression and secretion of specific hormones, which in turn control critical components of overall physiology. The basic function of these cells is understood; however, the molecular events involved in their hormonal regulation are not yet fully defined. While previously established cell lines have provided much insight into these regulatory mechanisms, the availability of representative cell lines from each cell lineage is limited, and currently none are derived from adult pituitary. We have therefore used retroviral transfer of SV40 T-antigen to mass immortalize primary pituitary cell culture from an adult mouse. We have generated 19 mixed cell cultures that contain cells from pituitary cell lineages, as determined by RT-PCR analysis and immunocytochemistry for specific hormones. Some lines expressed markers associated with multipotent adult progenitor cells or transit-amplifying cells, including SOX2, nestin, S100, and SOX9. The progenitor lines were exposed to an adenylate cyclase activator, forskolin, over 7 days and were induced to differentiate to a more mature gonadotrope cell, expressing significant levels of α-subunit, LHβ, and FSHβ mRNAs. Additionally, clonal populations of differentiated gonadotropes were exposed to 30 nM gonadotropin-releasing hormone and responded appropriately with a significant increase in α-subunit and LHβ transcription. Further, exposure of the lines to a pulse paradigm of GnRH, in combination with 17β-estradiol and dexamethasone, significantly increased GnRH receptor mRNA levels. This array of adult-derived pituitary cell models will be valuable for both studies of progenitor cell characteristics and modulation, and the molecular analysis of individual pituitary cell lineages

Dissecting the Transcriptional Regulatory Properties of Human Chromosome 16 Highly Conserved Non-Coding Regions

Non-coding DNA conservation across species has been often used as a predictor for transcriptional enhancer activity. However, only a few systematic analyses of the function of these highly conserved non-coding regions (HCNRs) have been performed. Here we use zebrafish transgenic assays to perform a systematic study of 113 HCNRs from human chromosome 16. By comparing transient and stable transgenesis, we show that the first method is highly inefficient, leading to 40% of false positives and 20% of false negatives. When analyzed in stable transgenic lines, a great majority of HCNRs were active in the central nervous system, although some of them drove expression in other organs such as the eye and the excretory system. Finally, by testing a fraction of the HCNRs lacking enhancer activity for in vivo insulator activity, we find that 20% of them may contain enhancer-blocking function. Altogether our data indicate that HCNRs may contain different types of cis-regulatory activity, including enhancer, insulators as well as other not yet discovered functions

Conservation planning in a fire-prone Mediterranean region: threats and opportunities for bird species

In response to the processes threatening biodiversity such as habitat loss, effective selection of priority conservation areas is required. However, reserve selection methods usually ignore the drivers of future habitat changes, thus compromising the effectiveness of conservation. In this work, we formulated an approach to explicitly quantify the impact of fire on conservation areas, considering such disturbance as a driver of land-cover changes. The estimated fire impact was integrated as a constraint in the reserve selection process to tackle the likely threats or opportunities that fire disturbance might cause to the targeted species depending on their habitat requirements. In this way, we selected conservation areas in a fire-prone Mediterranean region for two bird assemblages: forest and open-habitat species. Differences in conservation areas selected before and after integrating the impact of fire in the reserve selection process were assessed. Integration of fire impact for forest species moved preferences towards areas that were less prone to burn. However, a larger area was required to achieve the same conservation goals. Conversely, integration of fire impacts for open-habitat species shifted preferences towards conservation areas in locations where the persistence of their required habitat is more likely (i.e. shrublands). In other words, we prioritized the conservation of not only the current distribution of open-habitat birds, but also the disturbance process (i.e. fire) that favours their preferred habitat and distributions in the long term. Finally, this work emphasizes the need to consider the opposing potential impacts of wildfires on species for an effective conservation planning

Swimming physiology of European silver eels (Anguilla anguilla L.): energetic costs and effects on sexual maturation and reproduction

The European eel migrates 5,000–6,000 km to the Sargasso Sea to reproduce. Because they venture into the ocean in a pre-pubertal state and reproduce after swimming for months, a strong interaction between swimming and sexual maturation is expected. Many swimming trials have been performed in 22 swim tunnels to elucidate their performance and the impact on maturation. European eels are able to swim long distances at a cost of 10–12 mg fat/km which is 4–6 times more efficient than salmonids. The total energy costs of reproduction correspond to 67% of the fat stores. During long distance swimming, the body composition stays the same showing that energy consumption calculations cannot be based on fat alone but need to be compensated for protein oxidation. The optimal swimming speed is 0.61–0.67 m s−1, which is ~60% higher than the generally assumed cruise speed of 0.4 m s−1 and implies that female eels may reach the Sargasso Sea within 3.5 months instead of the assumed 6 months. Swimming trials showed lipid deposition and oocyte growth, which are the first steps of sexual maturation. To investigate effects of oceanic migration on maturation, we simulated group-wise migration in a large swim-gutter with seawater. These trials showed suppressed gonadotropin expression and vitellogenesis in females, while in contrast continued sexual maturation was observed in silver males. The induction of lipid deposition in the oocytes and the inhibition of vitellogenesis by swimming in females suggest a natural sequence of events quite different from artificial maturation protocols