Hormone homeostasis and induction of the small-fruit phenotype in ‘Hass’ avocado

Plant hormones play an important regulatory role in growth and development of fleshy fruits and often these are synthesized in a sequence that is tissue- and/or fruit-specific. Previous studies using Hass avocado and its small-fruit phenotype as a model demonstrated a relationship between the cytokinin/abscisic acid ration and final fruit size. More detailed studies revealed that differential activity of molybdenum-cofactor-containing enzymes was in part responsible for the observed increase in abscisic acid in small Hass fruit and that the same suite of enzymes likely impacted levels of indole-3-acetic acid. The present study confirms that the inability of the seed to produce/accumulate indole-3-acetic acid is a major contributing factor and characteristic of fruit that display a small-fruit phenotype, that reduced indole-3-acetic acid levels occur concomitant with increased abscisic acid, and that cytokinin metabolism appears unchanged irrespective of fruit size. Results are discussed in relation to the role of molybdenum-cofactor-requiring enzymes in indole-3-acetic acid and abscisic acid metabolism.     

Behavioural Reproductive Isolation in a Rotifer Hybrid Zone

A hybrid zone between two Brachionus plicatilis rotifer mitochondrial DNA (mtDNA) lineages was recently described in the Iberian Peninsula between a pond (Santed 2) and a lake (Gallocanta). The patterns of mitochondrial and nuclear genetic variation observed suggested that gene flow is mainly male-mediated from the lake to the pond. Here we test two hypotheses: (a) that male-mediated gene flow occurs through assortative mating between individuals from these ponds, (b) that behavioural isolation occurs between the two mtDNA lineages. We isolated, reared and genotyped rotifer clones from resting eggs collected in the sediments of these and two other distant ponds. We devised a quick, inexpensive RFLP method to discriminate between B. plicatilis and its sibling species B. ‘Manjavaeas’ and between both mtDNA B. plicatilis lineages. Behavioural no-choice tests using new-born, virgin males and females were performed between five clones. B. ‘Manjavacas’ and B. plicatilis were reproductively isolated. B. plicatilis clones did not show evidence of reproductive isolation, regardless of their mtDNA lineage, except Santed 2 males, which discriminated strongly against Gallocanta females. These results could help to explain the discrepancies between mitochondrial and nuclear genetic variation reported in the two populations.     

Saccharomyces cerevisiae-based molecular tool kit for manipulation of genes from gram-negative bacteria

A tool kit of vectors was designed to manipulate and express genes from a wide range of gram-negative species by using in vivo recombination. Saccharomyces cerevisiae can use its native recombination proteins to combine several amplicons in a single transformation step with high efficiency. We show that this technology is particularly useful for vector design. Shuttle, suicide, and expression vectors useful in a diverse group of bacteria are described and utilized. This report describes the use of these vectors to mutate clpX and clpP of the opportunistic pathogen Pseudomonas aeruginosa and to explore their roles in biofilm formation and surface motility. Complementation of the rhamnolipid biosynthetic gene rhlB is also described. Expression vectors are used for controlled expression of genes in two pseudomonad species. To demonstrate the facility of building complicated constructs with this technique, the recombination of four PCR-generated amplicons in a single step at >80% efficiency into one of these vectors is shown. These tools can be used for genetic studies of pseudomonads and many other gram-negative bacteria.     

Imprints of glacial refugia in the modern genetic diversity of Pinus sylvestris

Aim To understand the impact of glacial refugia and migration pathways on the modern genetic diversity of Pinus sylvestris. Location The study was carried out throughout Europe. Methods An extended set of data of pollen and macrofossil remains was used to locate the glacial refugia and reconstruct the migrating routes of P. sylvestris throughout Europe. A vegetation model was used to simulate the extent of the potential refugia during the last glacial period. At the same time a genetic survey was carried out on this species. Results The simulated distribution of P. sylvestris during the last glacial period is coherent with the observed fossil data, which showed a patchy distribution of the refugia between c. 40° N and 50° N. Several migrational fronts were detected within the Iberian and the Italian peninsulas, and outside the Hungarian plain and around the Alps. The modern mitochondrial DNA depicted three different haplotypes for P. sylvestris. Two distinct haplotypes were restricted to northern Spain and Italy, and the third haplotype dominated most of the present‐day remaining distribution range of P. sylvestris in Europe. Main conclusions During the last glacial period P. sylvestris was constrained under severe climatic conditions to survive in scattered and restricted refugial areas. Combining palaeoenvironmental data, vegetation modelling and the genetic data, we have shown that the long‐term isolation in the glacial refugia and the migrational process during the Holocene have played a major role in shaping the modern genetic diversity of P. sylvestris in Europe.     

BaySTDetect: detecting unusual temporal patterns in small area data via Bayesian model choice

Space-time modeling of small area data is often used in epidemiology for mapping chronic disease rates and by government statistical agencies for producing local estimates of, for example, unemployment or crime rates. Although there is typically a general temporal trend, which affects all areas similarly, abrupt changes may occur in a particular area, e.g. due to emergence of localized predictors/risk factor(s) or impact of a new policy. Detection of areas with "unusual" temporal patterns is therefore important as a screening tool for further investigations. In this paper, we propose BaySTDetect, a novel detection method for short-time series of small area data using Bayesian model choice between two competing space-time models. The first model is a multiplicative decomposition of the area effect and the temporal effect, assuming one common temporal pattern across the whole study region. The second model estimates the time trends independently for each area. For each area, the posterior probability of belonging to the common trend model is calculated, which is then used to classify the local time trend as unusual or not. Crucial to any detection method, we provide a Bayesian estimate of the false discovery rate (FDR). A comprehensive simulation study has demonstrated the consistent good performance of BaySTDetect in detecting various realistic departure patterns in addition to estimating well the FDR. The proposed method is applied retrospectively to mortality data on chronic obstructive pulmonary disease (COPD) in England and Wales between 1990 and 1997 (a) to test a hypothesis that a government policy increased the diagnosis of COPD and (b) to perform surveillance. While results showed no evidence supporting the hypothesis regarding the policy, an identified unusual district (Tower Hamlets in inner London) was later recognized to have higher than national rates of hospital readmission and mortality due to COPD by the National Health Service, which initiated various local enhanced services to tackle the problem. Our method would have led to an early detection of this local health issue.     

Spatial analysis of anthropogenic disturbance regimes and roadside shrubs in a fragmented agricultural landscape

Abstract. In fragmented woodlands in agricultural regions of Australia, roadside environments (road verges) provide important refuges for threatened native fauna and isolated populations of plant species. However, as roads are transport corridors for humans and their vehicles, species survival is affected through destruction, fragmentation and modification of remaining habitat by human activity. Few studies have recognised the effects of anthropogenic disturbances, in the form of historical roadworks activities, on adjacent roadside plant populations. The aim of this study was to investigate the effects of soil disturbance from roadworks on the spatial patterns and structural dynamics of roadside Acacia populations in an agricultural area in southeastern Australia. Stem size and disturbance data were collected from 135 roadside populations of three species of Acacia shrubs in the Lockhart Shire area. Spatial pattern analysis using the Network K‐function and Discriminant Function analyses showed that road verge width, road category, disturbance intensity and distance to nearest town were highly significant variables in relation to disturbance from roadworks and shrub structural dynamics. Colonising populations were more abundant along gravel roads where soil disturbance intensity was high, whereas stable populations were more abundant where soil disturbance intensity was low. Senescent populations were more abundant in narrow, little used road verge corridors 4–6 km from nearby towns. These results suggest that anthropogenic disturbance regimes may be critical to shrub recruitment and persistence in roadside environments, which has important consequences for conservation of threatened native flora and fauna in agricultural landscapes.     

Fruit size: Towards an understanding of the metabolic control of fruit growth using avocado as a model system

Final fruit size is the consequence of complex metabolic events that occur between fruit set and maturation. Disruption of these biochemical and molecular processes at any stage during fruit growth will impact on final fruit size. Because fruit size is a function of cell number rather than cell size, factors affecting cell division cycle activity assume importance. In this paper, we focus attention on the metabolic control of fruit growth using avocado as a model system. Three areas of current interest are highlighted, viz. the contribution by isoprenoid metabolism in the control of cell proliferation, the role played by carbohydrate content and composition in signalling changes in metabolite status and gene expression and maintenance of plant hormone homeostasis. Central to the process of fruit growth and control of final fruit size by cell division is 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and activity of the sucrose non-fermenting 1-related protein kinase (SnRK1) complex. It is argued that sugar content and composition of sink cells impact on SnRK1 (and hexokinase) to modulate expression of sugar-metabolizing enzymes, HMGR and molybdenum cofactor (MoCo)-containing enzymes. These changes, in turn, impact on hormone metabolism by affecting allocation of the purine-derived MoCo to aldehyde oxidase and thus the endogenous concentration of indole-3-acetic acid, abscisic acid and cytokinin (CK) to alter plant hormone homeostasis. These aspects are integrated into a model to explain the metabolic control of avocado fruit growth and final fruit size.     

The interplay between colonization history and gene flow in passively dispersing zooplankton: microsatellite analysis of rotifer resting egg banks

Zooplanktonic organisms that disperse passively as diapausing eggs often exhibit surprisingly strong population subdivision given their high colonization ability. Here we attempt to disentangle the impacts of colonization history and gene flow on these organisms by studying the population genetic structure of the rotifer Brachionus plicatilis. The resting egg banks of B. plicatilis in fourteen salt lake populations in the Iberian Peninsula were examined using seven microsatellite loci. A remarkably high degree of geographical structuring was found (F_st=0.43), with a significant pattern of isolation by distance. Microsatellite loci were in genetic equilibrium, ruling out inbreeding as an important force in population structuring. Comparisons are drawn with previously published phylogeographical data. Surprisingly, introgression of nuclear genes was detected in neighbouring populations with divergent mtDNA haplotypes. These results stress the long lasting impact of colonization history and the modulating effect of gene flow at local scales in these organisms.     

In Vivo Evidence for Serine Biosynthesis-Defined Sensitivity of Lung Metastasis,but Not of Primary Breast Tumors,to mTORC1 Inhibition

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