Factors affecting the colonisation of a root system by fluorescent Pseudomonads: The effects of water,temperature and soil microflora

The colonisation of a radish root system by strains ofPseudomonas fluorescens, selected for their ability to promote potato and radish growth under different environmental conditions is reported. In pot experiments colonisation of different parts of the root system was measured at different temperatures, in different watering regimes and in sterile and recropped soil. Root colonisation was extensive but populations were highest on the upper root system and their distribution throughout the root system was greatly affected by environmental factors. percolation of water through the soil and partial soil sterilisation enhanced colonisation but the effects of temperature and recropping were complex. Growth promotion was unpredictable and there was no simple relationship between PGPR colonisation and stimulation of plant growth.     

The management of rice straw,fertilisers and leaf litters in rice cropping systems in Northeast Thailand.

The maintenance of soil organic matter (SOM) and the balancing of nutrient flows into and out of the rainfed rice cropping systems in Northeast Thailand is of paramount importance to arresting the decline in soil fertility and crop yields. A system where small applications of leaf litters from locally grown trees are applied annually to rice paddy soils prior to transplanting is described. The annual application of 1500 kg/ha of Cajanus cajan, Acacia auriculiformis, Phyllanthus taxodifolius and Samanea saman for five seasons resulted in increases in rice grain yield of 48, 35, 32 and 23% above the no-leaf litter control, respectively. Average annual nutrient inputs from the leaf litters, in kg/ha, ranged from 62.7 N, 3.9 P, 17.9 K, and 3.5 S for Cajanus cajan to 24.5 N, 1.5 P, 8.1 K and 2.0 S for Acacia auriculiformis. Nutrient balances, determined by the difference between the inputs (fertiliser and added leaf litters) and outputs (grain and straw) indicated net positive N and P balances of up to 457 and 60 kg/ha. respectively, after five seasons of leaf litter applications. Sulfur and potassium balances resulted in net deficits of up to −3 and −52 kg S and K/ha, respectively, where no leaf litter was applied and rice straw was removed following harvest. Calculated apparent nutrient recoveries reflected the decomposition rate of the added residues and were highest for P and K, reflecting their higher soil residual value than mobile nutrients such as N and S. Sustainable farming systems will require that crop yields are stable through the maintenance of soil fertility and the balance of nutrients in the system. This revised version was published online in June 2006 with corrections to the Cover Date.     

In vitro studies of siderophore production by wild type and rifampicin resistant strains of fluorescent Pseudomonads

Wild type (Rif⁻) and rifampicin resistant (Rif⁺) isolates of fluorescent Pseudomonads were grown in King’s B liquid medium in the presence and absence of iron and subcultured every 48 h for 12 days. Growth rates and the proportion of non-fluorescent colonies were measured. All Rif⁺ isolates when grown in the presence of iron produced a significantly higher proportion of non-fluorescent colonies. One Rif⁺ isolate had a reduced growth rate.     

A comparison of methods for measuring the colonisation of a root system by fluorescent Pseudomonads

Methods are described for measuring the colonisation of a radish (Raphanus sativus) root system by seedlings with rifampicin resistant fluorescent pseudomonads by dilution plating, and which would take account of differences in root morphology. Differences in the levels of pseudomonad colonisation was highly dependent on the units in which surface areas was expressed. Population levels are expressed using estimates of surface area based on root length, tap-root length and root weight. The best estimate of surface area was root length, but the most practical method was surface area calculated as a function of dry weight. This method could differentiate differences in the levels of root colonisation independent of differences in root morphology and was efficient enough to allow the routine processing of a large number of replicate root samples.     

Differentiated function and localisation of SPO11-1 and PRD3 on the chromosome axis during meiotic DSB formation in Arabidopsis thaliana

During meiosis, DNA double-strand breaks (DSBs) occur throughout the genome, a subset of which are repaired to form reciprocal crossovers between chromosomes. Crossovers are essential to ensure balanced chromosome segregation and to create new combinations of genetic variation. Meiotic DSBs are formed by a topoisomerase-VI-like complex, containing catalytic (e.g. SPO11) proteins and auxiliary (e.g. PRD3) proteins. Meiotic DSBs are formed in chromatin loops tethered to a linear chromosome axis, but the interrelationship between DSB-promoting factors and the axis is not fully understood. Here, we study the localisation of SPO11-1 and PRD3 during meiosis, and investigate their respective functions in relation to the chromosome axis. Using immunocytogenetics, we observed that the localisation of SPO11-1 overlaps relatively weakly with the chromosome axis and RAD51, a marker of meiotic DSBs, and that SPO11-1 recruitment to chromatin is genetically independent of the axis. In contrast, PRD3 localisation correlates more strongly with RAD51 and the chromosome axis. This indicates that PRD3 likely forms a functional link between SPO11-1 and the chromosome axis to promote meiotic DSB formation. We also uncovered a new function of SPO11-1 in the nucleation of the synaptonemal complex protein ZYP1. We demonstrate that chromosome co-alignment associated with ZYP1 deposition can occur in the absence of DSBs, and is dependent on SPO11-1, but not PRD3. Lastly, we show that the progression of meiosis is influenced by the presence of aberrant chromosomal connections, but not by the absence of DSBs or synapsis. Altogether, our study provides mechanistic insights into the control of meiotic DSB formation and reveals diverse functional interactions between SPO11-1, PRD3 and the chromosome axis.     

N-glycosylation analysis of the human Tweety family of putative chloride ion channels supports a penta-spanning membrane arrangement: impact of N-glycosylation on cellular processing of Tweety homologue 2 (TTYH2)

The Tweety proteins are a family of recently identified putative Cl(-) channels predicted to be modified by N-glycosylation and, controversially, to contain five or six membrane-spanning domains, leading to the contentious proposal that members of this family do not share the same topology at the plasma membrane. In humans, three family members have been identified, designated TTYH1 (Tweety homologue 1), TTYH2 and TTYH3. To gain greater insight into the arrangement of membrane-spanning domains and cellular processing of Tweety proteins, in the present study we have examined the sequence homology, hydrophobicity and N-glycan content of members of this family and performed N-glycosylation site-mutagenesis studies on TTYH2 and TTYH3. Based on these observations we propose a structure for Tweety family proteins which incorporates five membrane-spanning domains with a topology at the cell surface in which the N-terminus is located extracellularly and the C-terminus cytoplasmically. Our results also suggest that N-glycosylation is important, but not essential, in the processing of members of the Tweety family with results indicating that, although incomplete N-glycosylation mediates reduced expression and increased ubiquitination of TTYH2, N-glycosylation is not the determining factor for TTYH2 trafficking to the plasma membrane. This information will be important for the characterization of Tweety family proteins in normal physiology and disease.     

Expression of the Intermediate Filament Keratin Gene,K15,in the Basal Cell Layers of Epithelia and the Hair Follicle

The intermediate filament keratin, K15, is present in variable abundance in stratified epithelia. In this study we have isolated and characterized the sheepK15gene, focusing on its expression in the follicles of sheep and mice. We show thatK15is expressed throughout the hair cycle in the basal layer of the outer root sheath that envelops the follicle. Strikingly, however, in large medullated wool follicles, a small group of basal outer root sheath cells located in the region thought to contain hair follicle stem cells areK15-negative. In the follicle bulbK15is expressed in cells situated next to the dermal papilla but not in the inner bulb cells. Elsewhere,K15is expressed at a low, variable level in the basal layer of the epidermis and sebaceous gland, often in a punctate pattern. In the esophagus of the sheepK15expression is restricted to the basal layer, in contrast to human esophagus where it is expressed throughout the epithelium. Transgenic mouse lines established with a 15-kb sheepK15gene construct exhibited faithful expression and showed no phenotypic consequences ofK15overexpression. An investigation of transgene expression showed thatK15is continuously expressed in outer root sheath cells during the hair cycle. Given its expression in the mitotically active basal cell layers of diverse epithelia and the follicle,K15expression appears to signal an early stage in the pathway of keratinocyte differentiation that precedes the decision of a cell to become epidermal or hair-like.