Identification and Characterisation CRN Effectors in Phytophthora capsici Shows Modularity and Functional Diversity

Phytophthora species secrete a large array of effectors during infection of their host plants. The Crinkler (CRN) gene family encodes a ubiquitous but understudied class of effectors with possible but as of yet unknown roles in infection. To appreciate CRN effector function in Phytophthora, we devised a simple Crn gene identification and annotation pipeline to improve effector prediction rates. We predicted 84 full-length CRN coding genes and assessed CRN effector domain diversity in sequenced Oomycete genomes. These analyses revealed evidence of CRN domain innovation in Phytophthora and expansion in the Peronosporales. We performed gene expression analyses to validate and define two classes of CRN effectors, each possibly contributing to infection at different stages. CRN localisation studies revealed that P. capsici CRN effector domains target the nucleus and accumulate in specific sub-nuclear compartments. Phenotypic analyses showed that few CRN domains induce necrosis when expressed in planta and that one cell death inducing effector, enhances P. capsici virulence on Nicotiana benthamiana. These results suggest that the CRN protein family form an important class of intracellular effectors that target the host nucleus during infection. These results combined with domain expansion in hemi-biotrophic and necrotrophic pathogens, suggests specific contributions to pathogen lifestyles. This work will bolster CRN identification efforts in other sequenced oomycete species and set the stage for future functional studies towards understanding CRN effector functions.     

Genetic analysis of the raffinose oligosaccharide pathway in lentil seeds

Two lentil (Lens culinaris) cultivars, Syrian Local Large (SLL) and PANT-L-406 (PL), have been used to study the genetics of the raffinose family of oligosaccharides (RFO) and a related compound, ciceritol, which is a galactosyl cyclitol. The RFO and ciceritol are the major soluble -galactosides in lentil seeds. Crosses were made between the two lentil lines and the patterns of inheritance for the total -galactoside content and for individual RFO compounds (raffinose, stachyose, verbascose) and ciceritol were determined in the embryos and seed coats from single seeds of the reciprocal F1s, the F2s and ten F3 families. The inheritance patterns for each of the -galactosides were complex and much of this complexity was attributed to an interaction between the embryo and its surrounding testa. A clear-cut segregation pattern was observed for verbascose. This was the result of PL embryos having very low, or no, verbascose. The F2 embryos had a segregation ratio of 3 high to 1 low level for this compound, suggesting that within this cross a low verbascose content was determined by a single recessive gene. There was good evidence from the F2 and F3 generations of a negative correlation between low levels of verbascose and high levels of ciceritol, which suggests a metabolic link between the RFO pathway and the pathway leading to ciceritol. The data are discussed in terms of defining strategies for genetically manipulating the-galactoside composition in lentil seeds, such that their negative effects on nutrition may be overcome without reducing significantly their positive role in abiotic stress resistance.     

The cover of living and dead corals from airborne remote sensing

Trends in coral cover are widely used to indicate the health of coral reefs but are costly to obtain from field survey over large areas. In situ studies of reflected spectra at the coral surface show that living and recently dead colonies can be distinguished. Here, we investigate whether such spectral differences can be detected using an airborne remote sensing instrument. The Compact Airborne Spectrographic Imager (Itres Research Ltd, Canada) was flown in two configurations: 10 spectral bands with 1-m² pixels and 6 spectral bands with 0.25-m² pixels. First, we show that an instrument with 10 spectral bands possesses adequate spectral resolution to distinguish living Porites, living Pocillopora spp., partially dead Porites, recently dead Porites (total colony mortality within 6 months), old dead (>6 months) Porites, Halimeda spp., and coralline red algae when there is no water column to confuse spectra. All substrata were distinguished using fourth-order spectral derivatives around 538 nm and 562 nm. Then, at a shallow site (Tivaru) at Rangiroa Atoll, Tuamotu Archipelago (French Polynesia), we show that live and dead coral can be distinguished from the air to a depth of at least 4 m using first- and fourth-order spectral derivatives between 562–580 nm. However, partially dead and recently dead Porites colonies could not be distinguished from an airborne platform. Spectral differences among substrata are then exploited to predict the cover of reef substrata in ten 25-m² plots at nearby Motu Nuhi (max depth 8 m). The actual cover in these plots was determined in situ using quadrats with a 0.01-m² grid. Considerable disparity occurred between field and image-based measures of substrate cover within individual 25-m² quadrats. At this small scale, disparity, measured as the absolute difference in cover between field and remote-sensing methods, reached 25% in some substrata but was always less than 10% for living coral (99% of which consisted of Porites spp.). At the scale of the reef (all ten 25-m² quadrats), however, disparities in percent cover between imagery and field data were less than 10% for all substrata and extremely low for some classes (e.g. <3% for living Porites, recently dead Porites and Halimeda). The least accurately estimated substrata were sand and coralline red algae, which were overestimated by absolute values 7.9% and 6.6%, respectively. The precision of sampling was similar for field and remote-sensing methods: field methods required 19 plots to detect a 10% difference in coral cover among three reefs with a statistical power of 95%. Remote-sensing methods required 21 plots. However, it took 1 h to acquire imagery over 92,500 m² of reef, which represents 3,700 plots of 25 m² each, compared with 3 days to survey 10 such plots underwater. There were no significant differences in accuracy between 1-m² and 0.25-m² image resolutions, suggesting that the advantage of using smaller pixels is offset by reduced spectral information and an increase in noise (noise was observed to be 1.6–1.8 times greater in 0.25-m² pixels). We show that airborne remote sensing can be used to monitor coral and algal cover over large areas, providing that water is shallow and clear, and that brown fleshy macroalgae are scarce, that depth is known independently (e.g. from sonar survey).     

Starch synthesis and carbon partitioning in developing endosperm

The biosynthesis of starch is the major determinant of yield in cereal grains. In this short review, attention is focused on the synthesis of the soluble substrate for starch synthesis, ADPglucose (ADPG). Consideration is given to the pathway of ADPG production, its subcellular compartmentation, and the role of metabolite transporters in mediating its delivery to the site of starch synthesis. As ADPG is an activated sugar, the dependence of its production on respiration, changes which occur during development, and the constraints which ATP production may place on carbon partitioning into different end-products are discussed.     

Atomic force microscopy of pea starch granules: granule architecture of wild-type parent, r and rb single mutants, and the rrb double mutant

AFM studies have been made of the internal structure of pea starch granules. The data obtained provides support for the blocklet model of starch granule structure (Carbohydr. Polym. 32 (1997) 177-191). The granules consist of hard blocklets dispersed in a softer matrix material. High-resolution images have yielded new insights into the detailed structure of growth rings within the granules. The blocklet structure is continuous throughout the granule and the growth rings originate from localised defects in blocklet production distributed around the surface of spheroidal shells within the granules. A mutation at the rb locus did not lead to significant changes in granule architecture. However, a mutation at the r locus led to loss of growth rings and changed blocklet structure. For this mutant the blocklets were distributed within a harder matrix material. This novel composite arrangement was used to explain why the granules had internal fissures and also changes in gelatinisation behaviour. It is suggested that the matrix material is the amylose component of the granule and that both amylose and amylopectin are present within the r mutant starch granules in a partially-crystalline form. Intermediate changes in granule architecture have been observed for the double mutant rrb.     

The effects of acute heat exposure on muscular strength,muscular endurance,and muscular power in the euhydrated athlete

The purpose of this study was to determine the effects of acute heat exposure upon muscular strength, muscular endurance, and muscular power in euhydrated athletes. Ten healthy, weight-trained men (average age = 23.0 +/- 4.0 years) volunteered for this investigation. Subjects were randomized to normothermic (22.5 degrees C, 65% relative humidity [RH]) or hyperthermic (65-75 degrees C, 15% RH) condition for 30 minutes. Results indicated that all subjects experienced significant (p < 0.05) hemodynamic stress because of the 30 minutes of heat exposure (blood pressure [BP](rest) 124/78 mm Hg to BP(postsauna) 148/60 mm Hg, heart rate [HR](rest) 64 b.min(-1) to HR(postsauna) 122 b.min(-1)). Oral and tympanic temperature measurements correlated strongly (r(2) = 0.904) and increased by 2.48 and 2.71 degrees C, respectively, during sauna exposure. One repetition maximum (1RM) bench press strength did not differ between the 2 conditions, whereas 1RM leg press strength was significantly decreased (p < 0.05) after the hyperthermic protocol. Subjects' muscular endurance decreased significantly (p < 0.05) in both the leg press (29.2%) and bench press (15.8%) after the sauna exposure. In contrast, muscular power (vertical jump) increased significantly (3.1%, p < 0.5) after acute heat exposure. In agreement with previous studies, we concluded that acute heat exposure is detrimental to muscular endurance; however, the areas of strength and power are far less unequivocal.