Establishment and characterization of India's first marine fish cell line (SISK) from the kidney of sea bass (Lates calcarifer)

A continuous cell line (SISK) from kidney of sea bass, Lates calcarifer, has been established and characterized. The cell line was maintained in Leibovitz' L-15 supplemented with 15% fetal bovine serum. This cell line has been subcultured more than 100 times over a period of 2 years. The SISK cell line consists of predominantly of epithelial-like cells. These cells showed strong positive for epithelial markers such as cytokeratin 19 and pancytokeratin. The cells were able to grow at temperature between 25 and 32 °C with optimum temperature of 28 °C. The growth rate of sea bass kidney cells increased as the FBS proportion increased from 2% to 20% at 28 °C with optimum growth at the concentrations of 15% or 20% FBS. The distribution of chromosome number was 30 to 56 with a modal peak at 48 chromosomes. Polymerase chain reaction products were obtained from SISK cells and tissues of sea bass with primer sets of microsatellite markers of sea bass. Five fish viruses were tested on this cell line to determine its susceptibility to these viruses and this was found to be susceptible to MABV NC1 and nodavirus, and the infection was confirmed by RT-PCR and CPE. This suggests that the SISK cell line has good potential for the isolation of various fish viruses. This cell line has been shown to be susceptible to bacterial extracellular products. The SISK cell line is the India's first marine fish cell line.

Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro

Development of continuous cell lines from shrimp is essential to investigate viral pathogens. Unfortunately, there is no valid cell line developed from crustaceans in general and shrimps in particular to address this issue. Lack of information on the requirements of cells in vitro limits the success of developing a cell line, where the microenvironment of a cell culture, provided by the growthmedium, is of prime importance. Screening and optimization of growth medium components based on statistical experimental designs have been widely used for improving the efficacy of cell culture media. Accordingly, we applied Plackett–Burman design and response surface methodology to study multifactorial interactions between the growth factors in shrimp cell culture medium and to identify the most important ones for growth of lymphoid cell culture from Penaeus monodon. The statistical screening and optimization indicated that insulin like growth factor-I (IGF-I) and insulin like growth factor-II (IGF-II) at concentrations of 100 and 150 ng ml-1, respectively, could significantly influence the metabolic activity and DNA synthesis of the lymphoid cells. An increase of 53 % metabolic activity and 24.8 % DNA synthesis could be obtained, which suggested that IGF-I and IGFII had critical roles in metabolic activity and DNA synthesis of shrimp lymphoid cells

Prion Protein is Expressed on Long-term Repopulating Hematopoietic Stem Cells and is Necessary for their Self-renewal

We show that the prion protein (PrP) is expressed on the surface of bone marrow cell populations enriched in long-term repopulating hematopoietic stem cells. Affinity purification of the PrP-positive and PrP-negative fractions from these populations, followed by competitive reconstitution assays, show that all long-term repopulating hematopoietic stem cells express PrP. Hematopoietic stem cells from PrP null bone marrow exhibit impaired self-renewal in serial competitive transplantation experiments, and premature exhaustion when exposed to cell cycle-specific myelotoxic injury. Therefore, PrP is a novel marker for hematopoietic stem cells and regulates their self-renewal.

AQA A2 Unit 5 Biology : control in cells and in organisms.

Guía de revisión en el área de la biología para estudiantes que estén preparando el examen AQA (Assessment and Qualifications Alliance) en el nivel AS (enseñanza secundaria de segundo ciclo). El libro está dividido en tres secciones: una introducción con información sobre el examen oficial, sugerencias, consejos y técnicas de estudio; una guía de contenido con los siguientes temas: cómo detectan y responden los organismos a los cambios en su medio ambiente, cómo coordinan sus respuestas a los estímulos, músculo esquelético, homeostasis, código genético, síntesis de proteínas y mutación genética, control de los genes y tecnología de clonación y sus aplicaciones; y un apartado final con ejemplos de preguntas y respuestas reales de los alumnos con comentarios del examinador.

Ex Vivo Construction of an Artificial Ocular Surface by Combination of Corneal Limbal Epithelial Cells and a Compressed Collagen Scaffold Containing Keratocytes

We have investigated the use of a laminin coated compressed collagen gel containing corneal fibroblasts (keratocytes) as a novel scaffold to support the growth of corneal limbal epithelial stem cells. The growth of limbal epithelial cells was compared between compressed collagen gel and a clinically proven conventional substrate, denuded amniotic membrane. Following compression of the collagen gel, encapsulated keratocytes remained viable and scanning electron microscopy showed that fibres within the compressed gel were dense, homogeneous and similar in structure to those within denuded amniotic membrane. Limbal epithelial cells were successfully expanded upon the compressed collagen resulting in stratified layers of cells containing desmosome and hemidesmosome structures. The resulting corneal constructs of both the groups shared a high degree of transparency, cell morphology and cell stratification. Similar protein expression profiles for cytokeratin 3 and cytokeratin 14 and no significant difference in cytokeratin 12 mRNA expression levels by real time PCR were also observed. This study provides the first line of evidence that a laminin coated compressed collagen gel containing keratocytes can adequately support limbal epithelial cell expansion, stratification and differentiation to a degree that is comparable to the leading conventional scaffold, denuded amniotic membrane.

Utilisation by upland rice of plant residue- and fertiliser-phosphorus in two tropical acid soils

A dual isotopic technique was used to assess the effects of soil type, and residues of Gliricidia sepium, without and with added fertiliser-P on the utilisation of P. Upland rice (Oryza sativa) was grown for 70 days in two tropical acid soils of different P sorbing capacity and P status. Uniformly P-32-labelled soils were treated with inorganic fertiliser-P tagged with P-33, Gliricidia sepium residue applied at planting and 3 weeks earlier, and in a combination of fertiliser-P and Gliricidia applied at and 3 weeks before planting. There were significant responses of shoot and root weights, and total P uptake to Gliricidia- and/or fertiliser-P addition in the Ultisol (low P status) but not the Oxisol (high P status), suggesting that P in the latter soil was not yield limiting, despite the high standard P requirement. Similarly, incorporation of Gliricidia three weeks before planting further increased shoot weight only in the Ultisol. There were generally higher proportions, quantities and percent utilisations of the Gliricidia- P and fertiliser-P in the Ultisol than in the Oxisol. Gliricidia significantly increased the utilisation of fertiliser-P only in the Ultisol. However, early application of Gliricidia increased Gliricidia- P but not fertiliser-P utilisation in the Ultisol. Added fertiliser-P did not influence Gliricidia- P utilisation.

Plant species and functional group effects on abiotic and microbial soil properties and plant-soil feedback responses in two grasslands

1 Plant species differ in their capacity to influence soil organic matter, soil nutrient availability and the composition of soil microbial communities. Their influences on soil properties result in net positive or negative feedback effects, which influence plant performance and plant community composition. 2 For two grassland systems, one on a sandy soil in the Netherlands and one on a chalk soil in the United Kingdom, we investigated how individual plant species grown in monocultures changed abiotic and biotic soil conditions. Then, we determined feedback effects of these soils to plants of the same or different species. Feedback effects were analysed at the level of plant species and plant taxonomic groups (grasses vs. forbs). 3 In the sandy soils, plant species differed in their effects on soil chemical properties, in particular potassium levels, but PLFA (phospholipid fatty acid) signatures of the soil microbial community did not differ between plant species. The effects of soil chemical properties were even greater when grasses and forbs were compared, especially because potassium levels were lower in grass monocultures. 4 In the chalk soil, there were no effects of plant species on soil chemical properties, but PLFA profiles differed significantly between soils from different monocultures. PLFA profiles differed between species, rather than between grasses and forbs. 5 In the feedback experiment, all plant species in sandy soils grew less vigorously in soils conditioned by grasses than in soils conditioned by forbs. These effects correlated significantly with soil chemical properties. None of the seven plant species showed significant differences between performance in soil conditioned by the same vs. other plant species. 6 In the chalk soil, Sanguisorba minor and in particular Briza media performed best in soil collected from conspecifics, while Bromus erectus performed best in soil from heterospecifics. There was no distinctive pattern between soils collected from forb and grass monocultures, and plant performance could not be related to soil chemical properties or PLFA signatures. 7 Our study shows that mechanisms of plant-soil feedback can depend on plant species, plant taxonomic (or functional) groups and site-specific differences in abiotic and biotic soil properties. Understanding how plant species can influence their rhizosphere, and how other plant species respond to these changes, will greatly enhance our understanding of the functioning and stability of ecosystems.

Effect of plant density and initial crown size on growth, development and yield in strawberry cultivars Elsanta and Bolero

The effects of density (plant spacing) and initial plant size on vegetative growth, flowering and fruiting were studied in the strawberry cultivars Elsanta and Bolero in their first and second years of cropping. The influence of these factors on light use and dry-matter partitioning was investigated. The size of planting material in 'Elsanta' and 'Bolero' slightly affected plant growth and yield, but this effect was not consistent and radiation use efficiency (RUE) and harvest index were unaltered. Plant spacing did not significantly affect the early stages of crop growth, but was important in determining growth and yield later in the season, this effect being more significant in the second year of cropping. Plant growth and yield per plant increased as plant spacing increased from 20 to 30 cm in both 'Elsanta' and 'Bolero', but the highest harvest index and yield per square metre were obtained at the closest spacing. Increased plant spacing also resulted in a greater leaf area and leaf area index. However, light was used less efficiently resulting in a lower RUE and lower harvest index (HI).

Plant species diversity, plant biomass and responses of the soil community on abandoned land across Europe: idiosyncracy or above-belowground time lags

The relationship between plant species diversity, productivity and the development of the soil community during early secondary succession on former arable land across Europe is investigated. The enhancement of biomass production due to the increase in initial plant species diversity and the consequent stimulation of soil microbial biomass and abundance of soil invertebrates are examined.

Vascular plant diversity and climate change in the alpine zone of the Lefka Ori, Crete

The aim of this study is to analyse the vascular flora and the local climate along an altitudinal gradient in the Lefka Ori massif Crete and to evaluate the potential effects of climate change on the plant diversity of the sub-alpine and alpine zones. It provides a quantitative/qualitative analysis of vegetation-environment relationships for four summits along an altitude gradient on the Lefka Ori massif Crete (1664-2339 m). The GLORIA multi-summit approach was used to provide vegetation and floristic data together with temperature records for every summit. Species richness and species turnover was calculated together with floristic similarity between the summits. 70 species were recorded, 20 of which were endemic, belonging to 23 different families. Cretan endemics dominate at these high altitudes. Species richness and turnover decreased with altitude. The two highest summits showed greater floristic similarity. Only 20% of the total flora recorded reaches the highest summit while 10% is common among summits. Overall there was a 4.96 degrees C decrease in temperature along the 675 m gradient. Given a scenario of temperature increase the ecotone between the sub-alpine and alpine zone would be likely to have the greatest species turnover. Southern exposures are likely to be invaded first by thermophilous species while northern exposures are likely to be more resistant to changes. Species distribution shifts will also depend on habitat availability. Many, already threatened, local endemic species will be affected first.

Regulated irrigation of woody ornamentals to improve plant quality and precondition against drought stress

Regulated irrigation has the potential to improve crop quality in woody ornamentals by reducing excessive vigour and promoting a more compact habit. This research aimed to compare the effectiveness and the mode of action of two techniques, regulated deficit irrigation (RDI) and partial root drying (PRD), when applied to container-grown ornamentals through drip irrigation. Results showed that RDI and PRD reduced growth in Cotinus coggygria 'Royal Purple', but in Forsythia x intermedia 'Lynwood', significant reductions were recorded only with RDI. Physiological measurements in Forsythia indicated that reductions in stomatal conductance (g(s)) occurred in both treatments, but those in the RDI tended to be more persistent. Reduced g(s) in PRD was consistent with the concept that chemical signals from the root can regulate stomatal aperture alone; however, the data also suggested that optimising the growth reduction required a moderate degree of shoot water deficit (i.e. a hydraulic signal to be imposed). As RDI was associated with tissue water deficit, it was used in a second experiment to determine the potential of this technique to precondition container-grown plants against subsequent drought stress (e.g. during retail stages or after planting out). Speed of acclimation would be important in a commercial context, and the results demonstrated that both slow and rapid imposition of RDI enabled Forsythia plants to acclimate against later drought events. This article discusses the potential to both improve ornamental plant quality and enhance tolerance to subsequent adverse conditions through controlled, regulated irrigation.

Plant chemistry and aphid parasitoids (Hymenoptera : Braconidae): Imprinting and memory

Emerging parasitoids of aphids encounter secondary plant chemistry from cues left by the mother parasitoid at oviposition and from the plant-feeding of the host aphid. In practice, however, it is secondary plant cheinistry oil the Surface of the aphid mummy which influences parasitoid olfactory behaviour. Offspring of Aphidius colemani reared oil Myzus persicae on artificial diet did no distinguish between the odours of bean and cabbage, but showed a clear preference for cabbage odour if sinigrin had been painted oil the back of the mummy. Similarly Aphidius rhopalosiphi reared on Metopolophium dirhodum on wheat preferred the odour of wheat plants grown near tomato plants to odour of wheat alone if the wheat plants oil which they had been reared had been exposed to the volatiles of nearby tomato plants. Aphidius rhopalosiphi reared on M dirhodum, and removed from the mummy before emergence, showed a preference for the odour of a different wheat cultivar if they had contacted a mummy from that cultivar, and similar results were obtained with A. colemani naturally emerged from M. persicae mummies. Aphidius colemani emerged from mummies oil one crucifer were allowed to contact in sequence (for 45 min each) mummies from two different crucifers. The mumber of attacks made in 10 min oil M. persicae was always significantly higher when aphids were feeding oil the same plant as the origin of the last MUMMY offered, or oil the second plant if aphids feeding on the third plant were not included. Chilling emerged A. colemani for 24 h at 5 degrees C appeared to erase the imprint of secondary plant chemistry, and they no longer showed host plant odour preferences in the olfactometer. When the parasitoids were chilled after three Successive mummy experiences, memory of the last experience appeared at least temporarily erased and preference was then shown for the chemistry of the second experience.

The effects of photoperiod and light spectrum on stock plant growth and rooting of cuttings of Cotinus coggygria 'Royal Purple'

Extending the season of production and improving the scheduling of ornamental crops are key commercial objectives for nurserymen. In some woody species, the period in which cuttings can be rooted successfully is transient, thus limiting the opportunities for scheduled production. Optimum rooting often occurs in early- to mid-summer coinciding with periods of active shoot growth. The relationship between this shoot activity and root initiation was investigated in Cotinus coggygria 'Royal Purple'. Shoot growth on stock plants was manipulated by altering the photoperiod or light quality. Results indicated there were seasonal effects on rooting, but the importance of shoot activity varied with harvest time. Cuttings harvested in August had high rooting percentages, irrespective of photoperiod, and despite shoot growth terminating in response to the short-day treatment. In contrast, by September, rooting percentage was highest in cuttings from plants under long-days, which had maintained greatest shoot growth activity. Cotinus shoots grown in vitro under 16 h days showed reduced shoot growth and increased rooting competence compared with shoots grown under 8 h days. Growing stock plants under polythene films, which altered the amount and quality of the incident light, influenced the rooting of cuttings harvested in August, but no consistent relationship with shoot activity was apparent. From a practical viewpoint, maintaining shoot activity late in the season may prolong the period for propagation by cuttings; but, from a scientific viewpoint, processes associated with an active shoot apex do not provide a complete explanation of seasonal variation in rooting.

Effect of plant density and initial crown size on growth, development and yield in strawberry cultivars Elsanta and Bolero

The effects of density (plant spacing) and initial plant size on vegetative growth, flowering and fruiting were studied in the strawberry cultivars Elsanta and Bolero in their first and second years of cropping. The influence of these factors on light use and dry-matter partitioning was investigated. The size of planting material in 'Elsanta' and 'Bolero' slightly affected plant growth and yield, but this effect was not consistent and radiation use efficiency (RUE) and harvest index were unaltered. Plant spacing did not significantly affect the early stages of crop growth, but was important in determining growth and yield later in the season, this effect being more significant in the second year of cropping. Plant growth and yield per plant increased as plant spacing increased from 20 to 30 cm in both 'Elsanta' and 'Bolero', but the highest harvest index and yield per square metre were obtained at the closest spacing. Increased plant spacing also resulted in a greater leaf area and leaf area index. However, light was used less efficiently resulting in a lower RUE and lower harvest index (HI).