A survey of Nicotiana germplasm for resistance to Tomato spotted wilt virus (TSWV)

The reaction to Tomato spotted wilt virus (TSWV) was evaluated in 94 accessions of Nicotiana, originating from the Institute of Soil Science and Plant Cultivation tobacco germplasm collection in Pu?awy, Poland. Tests for resistance were conducted under greenhouse conditions using single TSWV isolate collected from tobacco plantation in Lublin district, Poland. The presence of the virus was verified using DAS-ELISA. SCAR markers associated with TSWV resistance gene were applied. The members of the section Alatae, the genus Nicotiana: N. alata, N. forgetiana, and Nicotiana x sanderae as well as N. tabacum cultivars: ‘Polalta’ and ‘Wiktoria’ with the TSWV resistance gene introduced from N. alata, displayed the hypersensitive reaction (HR) against TSWV (grade 0 on symptom intensity scale). In some of those accessions, the virus spread from the initially infected areas eliciting systemic hypersensitive reaction (SHR). Five accessions of N. alata and three of Nicotiana x sanderae were composed of 6.3–50.0 % of plants in which SHR symptoms appeared. In all of N. forgetiana plants HR reaction was followed by systemic infection (SHR). In N. tabacum ‘Wiktoria’ 21.1 % of plants showed HR reaction, while the remaining were susceptible (S). All of the genotypes which responded with HR or SHR reaction to TSWV infection demonstrated the presence of SCAR markers linked to the resistance gene. The remaining eighty tested accessions were identified as being susceptible upon exposure to TSWV.     

Nitrogen transformations and pools in N-saturated mountain spruce forest soils

Nitrogen leaching persists in mountain forests of Europe even in the presence of decreasing N depositions. We have hypothesized that this leaching is linked to soil N transformations occurring over the whole year, even at 0°C temperatures. The aims were to estimate (1) the effect of temperature on N transformations and (2) N pools and fluxes. The study sites are situated in the Bohemian Forest (Czech Republic). Litter, humus, and 0–10-cm mineral layers were sampled in early spring, and the effect of temperature on net nitrification, net ammonification, and microbial N immobilization were measured in a short-term incubation experiment without substrate addition. Nitrogen pools were calculated from the concentrations of N forms in the soil and soil pool weights, while daily N fluxes were calculated from daily net rates of processes and soil pool weights. Relationships between temperature and net nitrification, net ammonification, and microbial N immobilization did not follow the Arrhenius type equation; all processes were active close to 0°C, indicating that microbial N transformations occur over the whole year. Microbial N immobilization rate was generally greater than N mineralization rate. The microbial N pool was significantly larger than mineral N pools. Organic layers containing tens of grams of available N per square meter contributed more than 70% to the available N in the soil profile. Daily N fluxes were related to N pools. On average, N fluxes represented daily mineral and microbial N pool changes of 1.14 and 1.95%, respectively. The effect of microbial composition on the C/N ratio of microbial biomass and respiration is discussed.     

Reproductive timing in three southern Appalachian stream fishes

We quantified the reproductive biology of mottled sculpin (Cottus bairdi), rosyside dace (Clinostomus funduloides) and longnose dace (Rhinichthus cataractae) in Coweeta Creek, North Carolina, USA, a fifth order stream. These species are abundant in coldwater streams through out the southern Appalachian mountains, the southernmost limit of their range. We collected fishes from Coweeta Creek every second month (between July 1984 and January 1986) and quantified the seasonal reproductive cycles of females, using both histological techniques and the relative gonadal index (RGI). Both techniques indicated that I) C. bairdi reproduced between March and May, 2) CI. funduloides spawned between May and August and 3) R. cataractae were reproductively active between June and July. These conclusions were supported by the following: 1) the presence of reproductive coloration, 2) gravid females, 3) courtship displays, 4) the presence of nest-guarding C. bairdi males and 5) the timing of young-of-the-year recruitment. The reproductive characteristics of these 3 species were correlated with different environmental variables. The mean monthly RGI values for both C. bairdi and R. cataractae were inversely correlated with both maximum and minimum daily water temperatures. The mean RGI values for C. bairdi also were negatively correlated with photoperiod, whereas the values for R. cataractae and CI. funduloides were positively correlated with the change in daily photo period.     

Analysis of leaf surface sesquiterpenes in potato varieties

A comparative study of potato leaf sesquiterpenes was carried out. GC, GC-MS, and NMR analyses were used to identify and quantify the sesquiterpenes in the leaf surfaces of 10 potato (Solanum tuberosum) varieties. Two sesquiterpene alcohols and 17 sesquiterpene hydrocarbons were identified and quantitatively determined. The distribution of the sesquiterpenes was found to be variety-specific. The sesquiterpene contents of the different potato varieties were subjected to cluster and principal component analyses. The eight potato varieties of the main chemotype cluster were dominated by beta-caryophyllene (9-148 ng/cm2), germacrene D (2-46 ng/cm2), germacrene D-4-ol (0.4-31 ng/cm2), beta-sesquiphellandrene (1-34 ng/cm2), and an unknown sesquiterpene alcohol III (0.2-37 ng/cm2). Chemometric classification distinguished two varieties, Mila and Vistula, from a major cluster. The Vistula variety was distinguished from the others by its high contents of beta-caryophyllene, alpha-humulene, germacrene D, and germacrene D-4-ol and the Mila variety by beta-elemene, trans-alpha-bergamotene, (Z)-beta-farnesene, (E)-beta-farnesene, trans-beta-bergamotene, beta-sesquiphellandrene, and unknown sesquiterpene alcohols I, II, III.     

The Hedgehog pathway promotes blood-brain barrier integrity and CNS immune quiescence

The blood-brain barrier (BBB) is composed of tightly bound endothelial cells (ECs) and perivascular astrocytes that regulate central nervous system (CNS) homeostasis. We showed that astrocytes secrete Sonic hedgehog and that BBB ECs express Hedgehog (Hh) receptors, which together promote BBB formation and integrity during embryonic development and adulthood. Using pharmacological inhibition and genetic inactivation of the Hh signaling pathway in ECs, we also demonstrated a critical role of the Hh pathway in promoting the immune quiescence of BBB ECs by decreasing the expression of proinflammatory mediators and the adhesion and migration of leukocytes, in vivo and in vitro. Overall, the Hh pathway provides a barrier-promoting effect and an endogenous anti-inflammatory balance to CNS-directed immune attacks, as occurs in multiple sclerosis.     

Plant immunity requires conformational changes [corrected] of NPR1 via S-nitrosylation and thioredoxins

Changes in redox status have been observed during immune responses in different organisms, but the associated signaling mechanisms are poorly understood. In plants, these redox changes regulate the conformation of NPR1, a master regulator of salicylic acid (SA)-mediated defense genes. NPR1 is sequestered in the cytoplasm as an oligomer through intermolecular disulfide bonds. We report that S-nitrosylation of NPR1 by S-nitrosoglutathione (GSNO) at cysteine-156 facilitates its oligomerization, which maintains protein homeostasis upon SA induction. Conversely, the SA-induced NPR1 oligomer-to-monomer reaction is catalyzed by thioredoxins (TRXs). Mutations in both NPR1 cysteine-156 and TRX compromised NPR1-mediated disease resistance. Thus, the regulation of NPR1 is through the opposing action of GSNO and TRX. These findings suggest a link between pathogen-triggered redox changes and gene regulation in plant immunity.     

Diversity of Ophiostomatales species associated with conifer-infesting beetles in the Western Carpathians

Bark beetles (Coleoptera: Curculionidae, Scolytinae) are commonly recognised as important agents of tree mortality in coniferous forests of the Western Carpathians. They, together with weevils, are consistently associated with ophiostomatoid fungi. Information regarding conifer beetle-associated fungi in the Western Carpathians remains incomplete and unreliable, particularly with respect to fir-infesting bark beetles. This study aims to clarify associations between fungi in the genera Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in Norway spruce (Picea abies), European larch (Larix decidua) and silver fir (Abies alba). Samples associated with 20 bark beetle species and weevils were collected from nine stands in Poland and the Czech Republic. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for four gene regions (ITS, LSU, ß-tubulin, TEF 1-α) and phylogenetic analyses. In total, 46 distinct taxa were identified, including 25 known and 21 currently unknown species. Several associations between fungi and subcortical insects were recorded for the first time. In addition, O. borealis and O. quercus were detected from A. alba for the first time. The composition of the fungal communities varied among the studied tree species and to a lesser degree among the beetle species. The spruce-infesting bark beetles were commonly associated with species of Leptographium s. l. and Ophiostoma s. str.; the larch-infesting bark beetles were often associated with Ophiostoma s. str. and Sporothrix, while the fir-infesting bark beetles were commonly associated with Ophiostoma s. str. and Graphilbum. The most commonly encountered fungal associates of the examined insects were (a) Grosmannia cucullata, G. piceiperda, Grosmannia sp. 1, Ophiostoma macroclavatum and O. piceae with the spruce-infesting bark beetles; (b) O. pseudocatenulatum and Sporothrix sp. 1 with the larch-infesting bark beetles; and (c) O. piceae, Ophiostoma sp. 2 and Graphilbum sp. 2 with the fir-infesting bark beetles. The differences in fungal associates among the bark beetle species occurring on P. abies, L. decidua and A. alba could be linked to the different habitats that these beetles occupy.