Physiological Responses of a Diazotrophic Cyanobacterium to Acidification of Paddy Floodwater: N2 Fixation, Photosynthesis, and Oxidative-Antioxidative Characteristics.

Long-term of excessive fertilization using nitrogen (N) chemical fertilizer caused the acidification of paddy soils. Presently, the impacts of soil acidification on physiological characteristics of diazotrophic cyanobacteria remain unknown. In order to elucidate this issue, the effects of paddy floodwater acidification on activities of respiration, photosynthetic oxygen evolution, and N2 fixation of a paddy diazotrophic cyanobacterium Aliinostoc sp. YYLX235 were investigated in this study. In addition, the origination and quenching of intracellular reactive oxygen species (ROS) were analyzed. The acidification of paddy floodwater decreased intracellular pH and interfered in energy flux from light-harvesting chlorophyll antenna to the reaction center of photosystem II (PS II). Activities of respiration, photosynthetic oxygen evolution, and N2 fixation were decreased by the acidification of paddy floodwater. Accompanied with an increase in ROS, the level of antioxidative system increased. Superoxide dismutase (SOD) and catalase (CAT) were the main enzymatic ROS scavengers in the cells of YYLX235; reduced glutathione (GSH) was the main non-enzymatic antioxidant. Antioxidants and oxidants in the cells of YYLX235 lost balance when the pH of paddy floodwater fell to 5.0 and 4.0, and lipid oxidative damage happened. The results presented in this study suggest that the acidification of paddy soil severely interfered in the photosynthesis of diazotrophic cyanobacteria and induced the production of ROS, which in turn resulted in oxidative damage on diazotrophic cyanobacteria and a decrease in cell vitality.

Duplication and transcriptional divergence of three Kunitz protease inhibitor genes that modulate insect and pathogen defenses in tea plant (Camellia sinensis).

Kunitz protease inhibitors (KPIs) are ubiquitous in plants and act as crucial compounds in defense responses against insect attack and pathogen infection. However, the influence of gene duplication on the postdivergence of the CsKPI genes involved in biotic stresses in tea plant is not well known. Here, we identified three CsKPI genes from tea plant (Camellia sinensis) and characterized their expression and evolutionary patterns among plant species. We found that CsKPI1, CsKPI2, and CsKPI3 diverged from their common ancestor 72.94 million years ago (MYA), and the tandem duplication of CsKPI2 and CsKPI3 occurred 26.78 MYA. An in vitro protein assay showed that the three CsKPI proteins were functional and inhibited the production of p-nitroanilide (PNA) from an artificial substrate. The three CsKPI-GFP fusion proteins localized to the cytoplasm. We showed that salicylic acid (SA) and transcripts of CsKPI2 and CsKPI3 significantly accumulated after infection with Glomerella cingulata. The application of exogenous SA stimulated the high expression of both CsKPI2 and CsKPI3 by activating cis-elements within their promoters. Under Ectropis oblique attack, CsKPI1 expression and jasmonic acid (JA) levels were more abundant in both insect-damaged leaf tissues and undamaged neighboring leaves. The application of jasmonic acid methyl ester elicited high expression levels of CsKPI1, suggesting that CsKPI1 accumulation requires JA production in tea plant. The overall findings suggest that the transcriptional divergence of KPI genes after duplication led to the specialized role of CsKPI1 in the physiological response to insect stress; the functional conservation between CsKPI2 and CsKPI3 confers resistance to pathogen infection in tea plant.

Transcriptomic and Phytochemical Analyses Reveal Root-Mediated Resource-Based Defense Response to Leaf Herbivory by Ectropis oblique in Tea Plant ( Camellia sinensis).

Leaf herbivory on tea plants ( Camellia sinensis) by tea geometrids ( Ectropis oblique) severely threaten the yield and quality of tea. In previous work, we found that local defense response was induced in damaged leaves by geometrids at the transcriptome level. Here, we investigated the systemic response triggered in undamaged roots and the potential role of roots in response to leaf herbivory. Comparative transcriptome analysis and carbohydrate dynamics indicated that leaf herbivory activated systemic carbon reallocation to enhance resource investment for local secondary metabolism. The crucial role of jasmonic acid and the involvement of other potential hormone signals for local and systemic signaling networks were supported by phytohormone quantification and dynamic expression analysis of phytohormone-related genes. This work represents a deep understanding of the interaction of tea plants and geometrids from the perspective of systems biology and reveals that tea plants have evolved an intricate root-mediated resource-based resistance strategy to cope with geometrid attack.

Growth inhibition and possible mechanism of oleamide against the toxin-producing cyanobacterium Microcystis aeruginosa NIES-843.

Oleamide, a fatty acid derivative, shows inhibitory effect against the bloom-forming cyanobacterium Microcystis aeruginosa. The EC50 of oleamide on the growth of M. aeruginosa NIES-843 was 8.60 ± 1.20 mg/L. In order to elucidate the possible mechanism of toxicity of oleamide against M. aeruginosa, chlorophyll fluorescence transient, cellular ultrastructure, fatty acids composition and the transcription of the mcyB gene involved in microcystins synthesis were studied. The results of chlorophyll fluorescence transient showed that oleamide could destruct the electron accepting side of the photosystem II of M. aeruginosa NIES-843. Cellular ultrastructure examination indicated that the destruction of fatty acid constituents, the distortion of thylakoid membrane and the loss of integrity of cell membrane were associated with oleamide treatment and concentration. The damage of cellular membrane increased the release of microcystins from intact cells into the medium. Results presented in this study provide new information on the possible mechanisms involved and potential utilization of oleamide as an algicide in cyanobacterial bloom control.

Silica fertilization and nano-MnO2 amendment on bacterial community composition in high arsenic paddy soils.

Silica fertilization and nano-MnO2 amendment are reported as useful approaches in lowering the accumulation of arsenic in rice grains, but the effects of silica fertilization or nano-MnO2 amendment on microbial community in the paddy soils containing high concentration of arsenic are still unknown. In order to elucidate this question, the structures and composition of microbial community in the paddy soils, in response to silica fertilization and nano-MnO2 amendment, were investigated using pyrosequencing technique. The results indicated that Proteobacteria, Chloroflexi, and Acidobacteria were the main dominating phyla in these paddy soils. A decrease in the relative abundance of Chloroflexi and Cyanobacteria, but an increase in the relative abundance of Acidobacteria was observed after silica fertilization and nano-MnO2 amendment. The changes of Acidobacteria, Chloroflexi, and Cyanobacteria were strongly correlated with pH and the concentration of bioavailable arsenic in the paddy soils. The α-diversity of bacteria in the paddy soils increased in response to silica fertilization at low amendment level, but decreased under silica or nano-MnO2 amendment at high amendment level. Results of ß-diversity analysis indicated that the microbial communities in the control treatment shared more similarity with that of those received low level of nano-MnO2 amendment, and the two silica fertilization treatments also shared more similarity with each other.