Role of Autophagy in Haematococcus lacustris Cell Growth under Salinity

The microalga Haematococcus lacustris (formerly H. pluvialis) is able to accumulate high amounts of the carotenoid astaxanthin in the course of adaptation to stresses like salinity. Technologies aimed at production of natural astaxanthin for commercial purposes often involve salinity stress; however, after a switch to stressful conditions, H. lacustris experiences massive cell death which negatively influences astaxanthin yield. This study addressed the possibility to improve cell survival in H. lacustris subjected to salinity via manipulation of the levels of autophagy using AZD8055, a known inhibitor of TOR kinase previously shown to accelerate autophagy in several microalgae. Addition of NaCl in concentrations of 0.2% or 0.8% to the growth medium induced formation of autophagosomes in H. lacustris, while simultaneous addition of AZD8055 up to a final concentration of 0.2 µM further stimulated this process. AZD8055 significantly improved the yield of H. lacustris cells after 5 days of exposure to 0.2% NaCl. Strikingly, this occurred by acceleration of cell growth, and not by acceleration of aplanospore formation. The level of astaxanthin synthesis was not affected by AZD8055. However, cytological data suggested a role of autophagosomes, lysosomes and Golgi cisternae in cell remodeling during high salt stress.

Characterization of Differentially Expressed Genes under Salt Stress in Olive

Climate change, currently taking place worldwide and also in the Mediterranean area, is leading to a reduction in water availability and to groundwater salinization. Olive represents one of the most efficient tree crops to face these scenarios, thanks to its natural ability to tolerate moderate salinity and drought. In the present work, four olive cultivars (Koroneiki, Picual, Royal de Cazorla and Fadak86) were exposed to high salt stress conditions (200 mM of NaCl) in greenhouse, in order to evaluate their tolerance level and to identify key genes involved in salt stress response. Molecular and physiological parameters, as well as plant growth and leaves’ ions Na+ and K+ content were measured. Results of the physiological measurements showed Royal de Cazorla as the most tolerant cultivar, and Fadak86 and Picual as the most susceptible ones. Ten candidate genes were analyzed and their complete genomic, CDS and protein sequences were identified. The expression analysis of their transcripts through reverse transcriptase quantitative PCR (RT-qPCR) demonstrated that only OeNHX7, OeP5CS, OeRD19A and OePetD were upregulated in tolerant cultivars, thus suggesting their key role in the activation of a salt tolerance mechanism.

Transcriptome Analysis of Arbuscular Mycorrhizal Casuarina glauca in Damage Mitigation of Roots on NaCl Stress

Casuarina glauca grows in coastal areas suffering long-term damage due to high salt stress. Arbuscular mycorrhizal fungi (AMF) can colonize their roots to alleviate the effects of salt stress. However, the specific molecular mechanism still needs to be further explored. Our physiological and biochemical analysis showed that Rhizophagus irregularis inoculation played an important role in promoting plant growth, regulating ion balance, and changing the activity of antioxidant enzymes. Transcriptome analysis of roots revealed that 1827 differentially expressed genes (DEGs) were affected by both R. irregularis inoculation and NaCl stress. The enrichment of GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) showed that most of these DEGs were significantly enriched in ion transport, antioxidant enzyme activity, carbohydrate metabolism, and cell wall. HAK5, KAT3, SKOR, PIP1-2, PER64, CPER, GLP10, MYB46, NAC43, WRKY1, and WRKY19 were speculated to play the important roles in the salt tolerance of C. glauca induced by R. irregularis. Our research systematically revealed the effect of R. irregularis on the gene expression of C. glauca roots under salt stress, laying a theoretical foundation for the future use of AMF to enhance plant tolerance to salt stress.

Salinity stress affects growth and physiology of mulberry (Morus sp.)

Mulberry (Morus sp.) plant is used to feed silkworms, and the leaves contain compounds with medicinal properties of secondary metabolites. However, the content of these compounds tends to increase under stress conditions, for instance, salt stress. This study, therefore, aimed to determine the accessions of mulberry with tolerance for salt stress. The stem cuttings of seven accessions from 5 regions, Bogor, Pati, Situbondo, Bali, and Gowa, were planted following a factorial randomized block design with 3 replications. Subsequently, the first factor using the accessions, and the second factor using NaCl solution (0.0%, 0.2%, 0.3%, and 0.4% concentrations) were performed. The variables observed were growth (leaves number, plant height, and shoots number), photosynthesis rate, total chlorophyll, and proline content. The results showed that the M6 accession exhibited tolerance under high salt stress, based on the leaves number, plant height, shoot number, photosynthesis rate, and proline content. Furthermore, an increase in salt concentration was discovered to cause a decrease in growth, photosynthesis rate, and total chlorophyll content. Also, proline accumulation stimulated by high salt stress possibly plays an important role in salinity tolerance.

Growth Performance Can Be Increased Under High Nitrate and High Salt Stress Through Enhanced Nitrate Reductase Activity in Arabidopsis Anthocyanin Over-Producing Mutant Plants

Nitrogen is one of the most important macro-nutrients for plant growth and crop productivity. The amount of synthetic nitrogen fertilizers supplied to crops has dramatically increased, leading to a notable rise in crop yields. However, excessive nitrogen use has an enormous negative impact on ecosystems and human health through the emission of intense greenhouse gases, such as nitric oxide derived from the nitrate (NO3–) assimilation cascade. Additionally, owing to the development of extensive irrigation in agriculture, crops are known to suffer from high salt stress. The effect of excessive nitrogen fertilizer application has been studied in some crops, but the effect of high nitrate level and salt stress on plant stress tolerance has not been studied in detail. Therefore, in this study we aimed to study the effects of high concentrations of NO3– on salt stress tolerance in Arabidopsis. In addition, since anthocyanin functions as a reactive oxygen species (ROS) scavenger under abiotic stress conditions, we investigated whether enhanced anthocyanin content helps Arabidopsis to withstand higher salt stress levels under high NO3– concentrations by using pap1-D/fls1ko double mutant plants, which accumulate excessive amount of anthocyanin. We found that Col-0 plants are more sensitive to salt stress under high NO3– concentrations. Although both the pap1-D/fls1ko and fls1ko plants accumulated higher anthocyanin levels and radical scavenging activities than Col-0 plants under both normal and salt stress conditions, the fls1ko plants exhibited much better growth than the pap1-D/fls1ko plants. It appears that the enhanced NR activities and transcript levels of NIA1 and NIA2 in pap1-D/fls1ko and fls1ko plants led to an increase in the synthesis of proteins and proline, which increases osmolytes against salt stress. Our results demonstrate that optimal levels of anthocyanin accumulation can enhance growth performance of plants under high NO3– and salt stress conditions.

Survival mechanism of a novel marine multistress-tolerant Meyerozyma guilliermondii GXDK6 under high NaCl stress as revealed by integrative omics analysis

A novel strain named Meyerozyma guilliermondii GXDK6 was provided in this work, which was confirmed to survive independently under high salt stress (12% NaCl) or co-stress condition of strong acid (pH 3.0) and high salts (10% NaCl) without sterilization. Its survival mechanism under high salt stress was revealed by integrated omics for the first time. Whole-genome analysis showed that 14 genes (e.g., GPD1 and FPS1) of GXDK6 relevant to salt tolerance were annotated and known to belong to various salt-resistant mechanisms (e.g., regulation of cell signal transduction and glycerol metabolism controls). Transcriptome sequencing results indicated that 1220 genes (accounting for 10.15%) of GXDK6 were differentially transcribed (p < 0.05) when GXDK6 growth was under 10% stress for 16 h, including important novel salt-tolerant-related genes (e.g., RTM1 and YHB1). Proteomics analysis demonstrated that 1005 proteins (accounting for 27.26%) of GXDK6 were differentially expressed (p < 0.05) when GXDK6 was stressed by 10% NaCl. Some of the differentially expressed proteins were defined as the novel salt-tolerant related proteins (e.g., sugar transporter STL1 and NADPH-dependent methylglyoxal reductase). Metabolomic analysis results showed that 63 types of metabolites (e.g., D-mannose, glycerol and inositol phosphate) of GXDK6 were up- or downregulated when stressed by 10% NaCl. Among them, D-mannose is one of the important metabolites that could enhance the salt-tolerance survival of GXDK6.

Multi-Omic Analyses Reveal Habitat Adaptation of Marine Cyanobacterium Synechocystis sp. PCC 7338

Cyanobacteria are considered as promising microbial cell factories producing a wide array of bio-products. Among them, Synechocystis sp. PCC 7338 has the advantage of growing in seawater, rather than requiring arable land or freshwater. Nonetheless, how this marine cyanobacterium grows under the high salt stress condition remains unknown. Here, we determined its complete genome sequence with the embedded regulatory elements and analyzed the transcriptional changes in response to a high-salt environment. Complete genome sequencing revealed a 3.70 mega base pair genome and three plasmids with a total of 3,589 genes annotated. Differential RNA-seq and Term-seq data aligned to the complete genome provided genome-wide information on genetic regulatory elements, including promoters, ribosome-binding sites, 5′- and 3′-untranslated regions, and terminators. Comparison with freshwater Synechocystis species revealed Synechocystis sp. PCC 7338 genome encodes additional genes, whose functions are related to ion channels to facilitate the adaptation to high salt and high osmotic pressure. Furthermore, a ferric uptake regulator binding motif was found in regulatory regions of various genes including SigF and the genes involved in energy metabolism, suggesting the iron-regulatory network is connected to not only the iron acquisition, but also response to high salt stress and photosynthesis. In addition, the transcriptomics analysis demonstrated a cyclic electron transport through photosystem I was actively used by the strain to satisfy the demand for ATP under high-salt environment. Our comprehensive analyses provide pivotal information to elucidate the genomic functions and regulations in Synechocystis sp. PCC 7338.

A high-throughput RNA-Seq approach to elucidate the transcriptional response of Piriformospora indica to high salt stress

Piriformospora indica, a root endophytic fungus, augments plant nutrition and productivity as well as protects plants against pathogens and abiotic stresses. High salinity is a major problem faced by plants as well as by microbes. Until now, the precise mechanism of salt stress tolerance in P. indica has remained elusive. In this study, the transcriptomes of control and salt-treated (0.5 M NaCl) P. indica were sequenced via the RNA-seq approach. A total of 30,567 transcripts and 15,410 unigenes for P. indica were obtained from 7.3 Gb clean reads. Overall 661 differentially expressed genes (DEGs) between control and treated samples were retrieved. Gene ontology (GO) and EuKaryotic Orthologous Groups (KOG) enrichments revealed that DEGs were specifically involved in metabolic and molecular processes, such as “response to salt stress”, “oxidoreductase activity”, “ADP binding”, “translation, ribosomal structure and biogenesis”, “cytoskeleton”, and others. The unigenes involved in “cell wall integrity”, “sterol biosynthesis”, and “oxidative stress” such as Rho-type GTPase, hydroxymethylglutaryl-CoA synthase, and thioredoxin peroxidase were up-regulated in P. indica subjected to salt stress. The salt-responsive DEGs have shown that they might have a potential role in salt stress regulation. Our study on the salt-responsive DEGs established a foundation for the elucidation of molecular mechanisms related to P. indica stress adaptation and a future reference for comparative functional genomics studies of biotechnologically important fungal species.

A genetic approach to dissect the role of prefoldins in Arabidopsis

SummaryThe prefoldin complex (PFDc) was identified in humans as co-chaperone of the cytosolic chaperonin TRiC/CCT. It is conserved in eukaryotes and is composed of subunits PFD1 to 6. PFDc-TRiC/CCT operates folding actin and tubulins. In addition to this function, PFDs participate in a wide range of cellular processes, both in the cytoplasm and in the nucleus, and their malfunction cause developmental alterations and disease in animals, and altered growth and environmental responses in yeast and plants. Genetic analyses in yeast indicate that not all functions performed by PFDs require the participation of the canonical complex. The lack of systematic genetic analyses in higher eukaryotes makes it difficult to discern whether PFDs participate in a particular process as canonical complex or in alternative configurations, i.e. as individual subunits or in other complexes. To tackle this question, and on the premise that the canonical complex cannot be formed if one subunit is missing, we have prepared an Arabidopsis mutant deficient in the six prefoldins, and compared various growth and environmental responses with those of the individual pfd. In this way, we demonstrate that the PFDc is required to delay flowering, for seed germination, or to respond to high salt stress, whereas two or more PFDs redundantly attenuate the response to osmotic stress. A coexpression analysis of differentially expressed genes in the sextuple mutant has identified several transcription factors, such as ABI5 or PIF4, acting downstream of PFDs. Furthermore, it has made possible to assign novel roles for PFDs, for instance, in the response to warm temperature.

Trade-off between shoot and root dry weight along with a steady CO2 assimilation rate ensures the survival of Eucalyptus camaldulensis under salt stress

Salt stress is a major challenge for reforestation in arid to semi-arid regions. Therefore the effect of salt stress was tested in 4-months-old saplings of Eucalyptus camaldulensis under controlled conditions. Individuals were subjected to three levels of salt stress (2, 8, 16 d·Sm^–1) and several traits describing growth and dry weight production/allocation, as well as physiological attributes were measured. The results showed that salt stress had no impact on plant height or stem diameter. Number of leaves, number of branches, and leaf chlorophyll content decreased significantly under high salt stress treatment. Leaf dry weight decreased significantly, but root dry weight increased significantly from 6.22 to 8.24 g under high salt stress treatment. Total plant dry weight remained similar while the root/shoot ratio increased significantly under high salt stress treatment. The net CO₂ assimilation rate remained stable at ~ 10.1 mmol·m^–2·s^–1 and stomatal conductance decreased significantly to 79 mmol·m^–2·s^–1 under high salt stress. Consequently, water use efficiency increased significantly to 3.25 mmol·mol^–1 under high salt stress. Therefore we may conclude that the young Eucalyptus camaldulensis saplings can tolerate moderate salt stress by increasing dry weight allocation towards the root system and sustaining the CO₂ assimilation rate.