LEA polypeptide profiling of recalcitrant and orthodox legume seeds reveals ABI3-regulated LEA protein abundance linked to desiccation tolerance

In contrast to orthodox seeds that acquire desiccation tolerance during maturation, recalcitrant seeds are unable to survive drying. These desiccation-sensitive seeds constitute an interesting model for comparative analysis with phylogenetically close species that are desiccation tolerant. Considering the importance of LEA (late embryogenesis abundant) proteins as protective molecules both in drought and in desiccation tolerance, the heat-stable proteome was characterized in cotyledons of the legume Castanospermum australe and it was compared with that of the orthodox model legume Medicago truncatula. RNA sequencing identified transcripts of 16 homologues out of 17 LEA genes for which polypeptides are detected in M. truncatula seeds. It is shown that for 12 LEA genes, polypeptides were either absent or strongly reduced in C. australe cotyledons compared with M. truncatula seeds. Instead, osmotically responsive, non-seed-specific dehydrins accumulated to high levels in the recalcitrant cotyledons compared with orthodox seeds. Next, M. truncatula mutants of the ABSCISIC ACID INSENSITIVE3 (ABI3) gene were characterized. Mature Mtabi3 seeds were found to be desiccation sensitive when dried below a critical water content of 0.4g H2O g DW–1. Characterization of the LEA proteome of the Mtabi3 seeds revealed a subset of LEA proteins with severely reduced abundance that were also found to be reduced or absent in C. australe cotyledons. Transcripts of these genes were indeed shown to be ABI3 responsive. The results highlight those LEA proteins that are critical to desiccation tolerance and suggest that comparable regulatory pathways responsible for their accumulation are missing in both desiccation-sensitive genotypes, revealing new insights into the mechanistic basis of the recalcitrant trait in seeds

Modelling and performance evaluation of storage enclosures

A model of moisture and heat transport was used to study the performance of storage enclosures. This paper examines several modelling approaches and presents the benefits and drawbacks of a ‘simple’ model which requires few input parameters. As a result, users do not need to measure many material properties, but some quality of the predictions is lost. The model is used to explore the balance of moisture exchange through ventilation holes and diffusion, the presence of buffering material inside enclosures and the effect of wall thickness. The predictions correspond well to experimental data measured in storage enclosures and a historic building. However, in order to bring modelling to the point where it can be used to engineer better enclosures, further research is needed. Experimental validation needs to be extensive and the limits of applicability of the model need to be clearly identified

Time Course Of Substance P Expression In Dorsal Root Ganglia Following Complete Spinal Nerve Transection

Recent evidence suggests that substance P (SP) is upregulated in primary sensory neurons following axotomy, and that this change occurs in larger neurons that do not usually produce SP. If so, this upregulation may allow normally neighboring, uninjured, and non-nociceptive dorsal root ganglion (DRG) neurons to become effective in activating pain pathways. Using immunohistochemistry, we performed a unilateral L5 spinal nerve transection upon male Wistar rats, and measured SP expression in ipsilateral L4 and L5 DRGs and contralateral L5 DRGs, at 1 to 14 days postoperatively (dpo), and in control and sham operated rats. In normal and sham operated DRGs, SP was detectable almost exclusively in small neurons (≤ 800 μm2). Following surgery, the mean size of SP-positive neurons from the axotomized L5 ganglia was greater at 2, 4, 7 and 14 dpo. Among large neurons (\u3e 800 μm2) from the axotomized L5, the percentage of SPpositive neurons increased at 2, 4, 7, and 14 dpo. Among small neurons from the axotomized L5, the percentage of SP-positive neurons was increased at 1 and 3 dpo, but was decreased at 7 and 14 dpo. Thus, SP expression is affected by axonal damage, and the time course of the expression is different between large and small DRG neurons. These data support a role of SP-producing, large DRG neurons in persistent sensory changes due to nerve injury

The effect of particulate matter on paper degradation

Background: In this work we explore the chemical effects of particulate matter on paper. We exposed paper made of pure cellulose to the environment in different locations in central London, outdoors (in sheltered conditions) and indoors, for a period of up to 6 months. We monitored particulate matter (PM) deposition by counting the particles deposited every month with a scanning electron microscope. We analysed elemental composition of the deposited particles using inductively coupled plasma mass spectrometry. After accelerated degradation of the exposed samples, we determined the degree of polymerisation using viscometry. Results: We observed higher deposition rates and higher metal concentration outdoors than indoors. Elemental analysis of the deposited particles revealed the presence of some transition metals (Fe, Cu, Cr) that can contribute to the degradation of cellulose fibres through the Fenton reaction. By comparing the degree of polymerisation of pro‑ tected, unprotected and unexposed samples we could determine the relative contribution of PM deposition on the increase of the degradation rate. We found that the surface concentration of iron correlates with the reduction in the degree of polymerisation of the exposed paper. Conclusions: The results suggest that the presence of Fenton metals in PM has a significant effect on the accelera‑ tion of the degradation of cellulose. However, we estimate that this will unlikely occur at the levels of area coverage by PM that are typically avoided in indoor heritage through preventive maintenance and cleaning