Importance of native arbuscular mycorrhizal inoculation in the halophyte Asteriscus maritimus for successful establishment and growth under saline conditions

Background and aims

The biological restoration of saline habitats could be achieved by using halophyte plant species together with adapted arbuscular mycorrhizal fungi (AMF). An interesting plant to be used in restoration of saline environments, Asteriscus maritimus, is highly mycotrophic. The aim of this study was to assess the effectiveness of native and allochthonous AMF to enhance the establishment and growth of the halophyte A. maritimus under saline conditions.

Methods

We studied the symbiotic effectiveness of four AMF strains (three native fungal isolates from a saline soil and one allochthonous, from collection) in A. maritimus subjected to increasing salinity stress. We measured plant physiological parameters by which AMF may ameliorate the detrimental effects of salinity stress.

Results

A. maritimus plants showed a high mycorrhizal dependency, even in absence of salt stress. Plants inoculated with native AMF had higher shoot dry weight, efficiency of photosystem II, stomatal conductance and accumulation of glutathione than those inoculated with the collection AMF at the highest level of salinity. Moreover, at this salt level, only 30 % of A. maritimus plants inoculated with the collection AMF survived, while with the three native AMF, the rate of survival was 100 %.

Conclusions

Results points out the importance of native AMF inoculation in the establishment, survival and growth of A. maritimus plants. Inoculation with these native AMF enhanced A. maritimus salt tolerance by increasing efficiency of photosystem II, stomatal conductance and glutathione content and by reducing oxidative damage. Thus, the use of adequate native AMF inocula could be a critical issue for success in recovering saline degraded areas.     

Characterization of salt-tolerance mechanisms in mycorrhizal (Claroideoglomus etunicatum) halophytic grass,Puccinellia distans

Responses of Puccinellia distans, a halophytic grass to low (50 mM) and high (200 mM) NaCl salinity, were studied in a sand culture experiment without or with inoculation by arbuscular mycorrhizal fungus (AMF), Claroideoglomus etunicatum isolated from its saline habitat. Plant biomass was not influenced by salinity levels, while a tendency to a higher biomass was observed in AMF plants under both control and saline conditions. Leaf photosynthesis increased by both salinity and AMF inoculation. Despite higher transpiration rate, AMF plants had higher water-use efficiency under sever saline conditions. AMF inoculation decreased proline concentration, but increased significantly leaf osmotic potential. Antioxidative enzymes responded differently to the salt and AMF treatments depending on the salt concentration and plant organ. Nonetheless, salt-induced malondialdehyde accumulation in the leaves diminished by AMF colonization. K and Ca contents were not affected by salt, while fungal colonization increased K in the roots and Ca in both leaves and roots. Our results indicated that enhancement of photosynthesis and ion homeostasis is involved in the tolerance of P. distans to both low and high salinity. AMF inoculation increased plants’ tolerance by augmentation of the above mechanisms accompanied by improvement of water relations and protection against oxidative damage in the leaves.     

Differential effects of abiotic factors and host plant traits on diversity and community composition of root-colonizing arbuscular mycorrhizal fungi in a salt-stressed ecosystem

Arbuscular mycorrhizal fungi (AMF) were investigated in roots of 18 host plant species in a salinized south coastal plain of Laizhou Bay, China. From 18 clone libraries of 18S rRNA genes, all of the 22 AMF phylotypes were identified into Glomus, of which 18 and 4 were classified in group A and B in the phylogenetic tree, respectively. The phylotypes related to morphologically defined Glomus species occurred generally in soil with higher salinity. AMF phylotype richness, Shannon index, and evenness were not significantly different between root samples from halophytes vs. non-halophytes, invades vs. natives, or annuals vs. perennials. However, AMF diversity estimates frequently differed along the saline gradient or among locations, but not among pH gradients. Moreover, UniFrac tests showed that both plant traits (salt tolerance, life style or origin) and abiotic factors (salinity, pH, or location) significantly affected the community composition of AMF colonizers. Redundancy and variation partitioning analyses revealed that soil salinity and pH, which respectively explained 6.9 and 4.2 % of the variation, were the most influential abiotic variables in shaping the AMF community structure. The presented data indicate that salt tolerance, life style, and origin traits of host species may not significantly affect the AMF diversity in roots, but do influence the community composition in this salinized ecosystem. The findings also highlight the importance of soil salinity and pH in driving the distribution of AMF in plant and soil systems.     

Arbuscular mycorrhizal symbiosis regulates the physiological responses,ion distribution and relevant gene expression to trigger salt stress tolerance in pistachio

Mycorrhizal symbiosis is generally considered effective in ameliorating plant tolerance to abiotic stress by altering gene expression, and evaluation of genes involved in ion homeostasis and nutrient uptake. This study aimed to use arbuscular mycorrhizal fungus (AMF) to alleviate salinity stress and analyse relevant gene expression in pistachio plants under No/NaCl stress in greenhouse conditions. Arbuscular mycorrhizal symbiosis was used to study the physiological responses, ion distribution and relevant gene expression in pistachio plants under salinity stress. After four months of symbiosis, mycorrhizal root colonization showed a significant reduction in all tested parameters under salt stress treatment compared to non-saline treatment. Salinity affected the morphological traits, and decreased the nutrient content including N, P, Mg and Fe as well as K/Na and Ca/Na ratios, relative water content (RWC), membrane stability index (MSI), and increased the concentration of K, Ca and Na nutrient, glycine betaine, ROS and MDA. Inoculation of seedlings with AMF mitigated the negative effects of salinity on plant growth as indicated by increasing the root colonization, morphological traits, glycine betaine, RWC and MSI. Specifically, under salinity stress, shoot and root dry weight, P and Fe nutrient content, K/Na and Ca/Na ratio of AMF plants were increased by 53.2, 48.6, 71.6, 60.2, 87.5, and 80.1% respectively, in contrast to those of the NMF plants. The contents of Na, O^2•− and MDA in AMF plants were significantly decreased by 66.8, 36.8, and 23.1%, respectively at 250 mM NaCl. Moreover, salinity markedly increased SOS1, CCX2 and SKOR genes expression and the inoculation with AMF modulated these genes expression; however, NRT2.4, PHO1 and PIP2.4 gene expressions were increased by salinity and AMF. It could be concluded that inoculation of AMF with Rhizophagus irregularis conferred a larger endurance towards soil salinity in pistachio plants and stimulate the nutrient uptake and ionic homeostasis maintenance, superior RWC and osmoprotection, toxic ion partitioning, maintaining membrane integrity and the ion-relevant genes expression.     

A meta-analysis of arbuscular mycorrhizal effects on plants grown under salt stress

Salt stress limits crop yield and sustainable agriculture in most arid and semiarid regions of the world. Arbuscular mycorrhizal fungi (AMF) are considered bio-ameliorators of soil salinity tolerance in plants. In evaluating AMF as significant predictors of mycorrhizal ecology, precise quantifiable changes in plant biomass and nutrient uptake under salt stress are crucial factors. Therefore, the objective of the present study was to analyze the magnitude of the effects of AMF inoculation on growth and nutrient uptake of plants under salt stress through meta-analyses. For this, data were compared in the context of mycorrhizal host plant species, plant family and functional group, herbaceous vs. woody plants, annual vs. perennial plants, and the level of salinity across 43 studies. Results indicate that, under saline conditions, AMF inoculation significantly increased total, shoot, and root biomass as well as phosphorous (P), nitrogen (N), and potassium (K) uptake. Activities of the antioxidant enzymes superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase also increased significantly in mycorrhizal compared to nonmycorrhizal plants growing under salt stress. In addition, sodium (Na) uptake decreased significantly in mycorrhizal plants, while changes in proline accumulation were not significant. Across most subsets of the data analysis, identities of AMF (Glomus fasciculatum) and host plants (Acacia nilotica, herbs, woody and perennial) were found to be essential in understanding plant responses to salinity stress. For the analyzed dataset, it is concluded that under salt stress, mycorrhizal plants have extensive root traits and mycorrhizal morphological traits which help the uptake of more P and K, together with the enhanced production of antioxidant enzymes resulting in salt stress alleviation and increased plant biomass.     

Arbuscular mycorrhiza and salt tolerance of plants

Although salt has detrimental effects on spore germination of arbuscular mycorrhizal fungi (AMF), their hyphal growth and the colonization rate of plants under laboratory conditions, many salt tolerant plants (the halophytes) are strongly colonized by AMF in their natural habitats. AMF spores in several saline soils consist of to up to 80 % of one single species, Glomus geosporum. In contrast, roots of halophytes are mostly colonized by fungi of the Glomus intraradices group, of which many are as yet uncultured. Salt stress is intimately related to drought in saline habitats. Molecular analyses of genes expressed upon salt stress indicate that aquaporins which facilitate the transfer of water across membranes play a major role in alleviating salt stress in plants. In AMF, genes serving to scavenge reactive oxygen species (ROS) are expressed upon exposure to salt, indicating that fungi have to develop an enhanced oxidative defence. The development of AMF inocula that confer sustained salt tolerance to plants would have enormous practical applications. Many positive reports on salt stress alleviation by AMF exist. However, the state of the art has not yet reached field applications. In contrast to other recent reviews, the present article focuses on ecological aspects of the symbiosis between AMF and halophytes. It also emphasizes the complexity of the interactions between salt and drought stress as well as the role of AMF in alleviating salt stress.     

Identification of Reference Genes for Quantitative Real-Time PCR in Date Palm (Phoenix dactylifera L.) Subjected to Drought and Salinity

Date palm is an important crop plant in the arid and semi-arid regions supporting human population in the Middle East and North Africa. These areas have been largely affected by drought and salinity due to insufficient rainfall and improper irrigation practices. Date palm is a relatively salt- and drought-tolerant plant and more recently efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Quantitative real-time PCR (qPCR) is a promising technique for the analysis of stress-induced differential gene expression, which involves the use of stable reference genes for normalizing gene expression. In an attempt to find the best reference genes for date palm’s drought and salinity research, we evaluated the stability of 12 most commonly used reference genes using the geNorm, NormFinder, BestKeeper statistical algorithms and the comparative ΔC_T method. The comprehensive results revealed that HEAT SHOCK PROTEIN (HSP), UBIQUITIN (UBQ) and YTH domain-containing family protein (YT521) were stable in drought-stressed leaves whereas GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (GAPDH), ACTIN and TUBULIN were stable in drought-stressed roots. On the other hand, SMALL SUBUNIT RIBOSOMAL RNA (25S), YT521 and 18S ribosomal RNA (18S); and UBQ, ACTIN and ELONGATION FACTOR 1-ALPHA (eEF1a) were stable in leaves and roots, respectively, under salt stress. The stability of these reference genes was verified by using the abiotic stress-responsive CYTOSOLIC Cu/Zn SUPEROXIDE DISMUTASE (Cyt-Cu/Zn SOD), an ABA RECEPTOR, and a PROLINE TRANSPORTER 2 (PRO) genes. A combination of top 2 or 3 stable reference genes were found to be suitable for normalization of the target gene expression and will facilitate gene expression analysis studies aimed at identifying functional genes associated with drought and salinity tolerance in date palm.     

Arbuscular mycorrhizal fungi native from a Mediterranean saline area enhance maize tolerance to salinity through improved ion homeostasis

Soil salinity restricts plant growth and productivity. Na⁺ represents the major ion causing toxicity because it competes with K⁺ for binding sites at the plasma membrane. Inoculation with arbuscular mycorrhizal fungi (AMF) can alleviate salt stress in the host plant through several mechanisms. These may include ion selection during the fungal uptake of nutrients from the soil or during transfer to the host plant. AM benefits could be enhanced when native AMF isolates are used. Thus, we investigated whether native AMF isolated from an area with problems of salinity and desertification can help maize plants to overcome the negative effects of salinity stress better than non‐AM plants or plants inoculated with non‐native AMF. Results showed that plants inoculated with two out the three native AMF had the highest shoot dry biomass at all salinity levels. Plants inoculated with the three native AMF showed significant increase of K⁺ and reduced Na⁺ accumulation as compared to non‐mycorrhizal plants, concomitantly with higher K⁺/Na⁺ ratios in their tissues. For the first time, these effects have been correlated with regulation of ZmAKT2, ZmSOS1 and ZmSKOR genes expression in the roots of maize, contributing to K⁺ and Na⁺ homeostasis in plants colonized by native AMF.     

Colonization with arbuscular mycorrhizal fungi improves salinity tolerance of tomato (Solanum lycopersicum L.) plants

The purpose of this study was to investigate the mechanisms underlying alleviation of salt stress by mycorrhization. Solanum lycopersicum L. cultivars Behta and Piazar with different salinity tolerance were cultivated in soil without salt (EC?=?0.63 dSm^?1), with low (EC?=?5 dSm^?1), or high (EC?=?10 dSm^?1) salinity. Plants inoculated with the arbuscular mycorrhizal fungi Glomus intraradices (+AMF) were compared to non-inoculated plants (?AMF). Under salinity, AMF-mediated growth stimulation was higher in more salt tolerant Piazar than in sensitive Behta. Mycorrhization alleviated salt-induced reduction of P, Ca, and K uptake. Ca/Na and K/Na ratios were also better in +AMF. However, growth improvement by AMF was independent from plant P nutrition under high salinity. Mycorrhization improved the net assimilation rates through both elevating stomatal conductance and protecting photochemical processes of PSII against salinity. Higher activity of ROS scavenging enzymes was concomitant with lowering of H₂O_2, less lipid peroxidation, and higher proline in +AMF. Cultivar differences in growth responses to salinity and mycorrhization could be well explained by differences in ion balance, photochemistry, and gas exchange of leaves. Function of antioxidant defenses seemed responsible for different AMF-responsiveness of cultivars under salinity. In conclusion, AMF may protect plants against salinity by alleviating the salt-induced oxidative stress.     

Effects of salt stress on nitrogenase activity and growth of four legumes

The effects of salt stress on nitrogenase (N2-ase) activity, growth and nitrogen content ofVicia faba (L.),Medicago sativa (L.) Merrill,Glycine max andVigna sinensis (L.) were investigated. Four levels of salinity were applied and salt treatments were imposed on inoculated and N-fertilized plants.M. sativa tolerated mild levels of salinity but higher salt concentrations depressed N₂-ase activity of this species. The other three legumes were considerably affected by salt treatments, and N₂-ase activity was significantly reduced by salinity. Vicia faba, carrying elongate nodules, could restore a partial N₂-ase activity upon recovery from salt stress whereasG. max andV. sinesis, both with spherical nodules, could not regain significant activity when salinity was removed. Salt stress retarded growth of both inoculated and N-fertilized plants. The nitrogen content of both treatments was also affected by salinity and the effect was more severe for inoculated than N-fertilized plants.     

Spindle assembly checkpoint genes reveal distinct as well as overlapping expression that implicates MDF-2/Mad2 in postembryonic seam cell proliferation in Caenorhabditis elegans

Background

The spindle assembly checkpoint (SAC) delays anaphase onset by inhibiting the activity of the anaphase promoting complex/cyclosome (APC/C) until all of the kinetochores have properly attached to the spindle. The importance of SAC genes for genome stability is well established; however, the roles these genes play, during postembryonic development of a multicellular organism, remain largely unexplored.

Results

We have used GFP fusions of 5' upstream intergenic regulatory sequences to assay spatiotemporal expression patterns of eight conserved genes implicated in the spindle assembly checkpoint function in Caenorhabditis elegans. We have shown that regulatory sequences for all of the SAC genes drive ubiquitous GFP expression during early embryonic development. However, postembryonic spatial analysis revealed distinct, tissue-specific expression of SAC genes with striking co-expression in seam cells, as well as in the gut. Additionally, we show that the absence of MDF-2/Mad2 (one of the checkpoint genes) leads to aberrant number and alignment of seam cell nuclei, defects mainly attributed to abnormal postembryonic cell proliferation. Furthermore, we show that these defects are completely rescued by fzy-1(h1983)/CDC20, suggesting that regulation of the APC/C^CDC20 by the SAC component MDF-2 is important for proper postembryonic cell proliferation.

Conclusion

Our results indicate that SAC genes display different tissue-specific expression patterns during postembryonic development in C. elegans with significant co-expression in hypodermal seam cells and gut cells, suggesting that these genes have distinct as well as overlapping roles in postembryonic development that may or may not be related to their established roles in mitosis. Furthermore, we provide evidence, by monitoring seam cell lineage, that one of the checkpoint genes is required for proper postembryonic cell proliferation. Importantly, our research provides the first evidence that postembryonic cell division is more sensitive to SAC loss, in particular MDF-2 loss, than embryonic cell division.     

Halotolerant Aerobic Heterotrophic Bacteria from the Great Salt Plains of Oklahoma

The Salt Plains National Wildlife Refuge (SPNWR) near Cherokee, Oklahoma, contains a barren salt flat where Permian brine rises to the surface and evaporates under dry conditions to leave a crust of white salt. Rainfall events dissolve the salt crust and create ephemeral streams and ponds. The rapidly changing salinity and high surface temperatures, salinity, and UV exposure make this an extreme environment. The Salt Plains Microbial Observatory (SPMO) examined the soil microbial community of this habitat using classic enrichment and isolation techniques and phylogenetic rDNA studies. Rich growth media have been emphasized that differ in total salt concentration and composition. Aerobic heterotrophic enrichments were performed under a variety of conditions. Heterotrophic enrichments and dilution plates have generated 105 bacterial isolates, representing 46 phylotypes. The bacterial isolates have been characterized phenotypically and subjected to rDNA sequencing and phylogenetic analyses. Fast-growing isolates obtained from enrichments with 10% salt are predominantly from the gamma subgroup of the Proteobacteria and from the low GC Gram-positive cluster. Several different areas on the salt flats have yielded a variety of isolates from the Gram-negative genera Halomonas, Idiomarina, Salinivibrio, and Bacteroidetes. Gram-positive bacteria are well represented in the culture collection including members of the Bacillus, Salibacillus, Oceanobacillus, and Halobacillus.     

Arbuscular mycorrhizal fungi enhances salinity tolerance of Panicum turgidum Forssk by altering photosynthetic and antioxidant pathways

Present experiments were conducted to assess the response of Panicum turgidum to salinity and possible role of arbuscular mycorrhizal fungi (AMF) in enhancing the salt tolerance. The activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione reductase (GR) and compatible solutes were increased by salt stress and were further enhanced by AMF inoculation. Hydrogen peroxide and malonaldehyde content increased in salt-stressed plants while a reduction was observed due to AMF inoculation. Salt-stressed plants showed higher activities of pyruvate orthophosphate dikinase (PPDK), phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase as compared to control and AMF-inoculated plants. Salt stress caused significant decrease in phosphorous, potassium and calcium uptake but an increase in sodium uptake was observed. AMF alleviate salinity-induced negative impact on the plant growth and nutrient uptake by reducing the oxidative damage through strengthening of the antioxidant system.     

Evaluation and selection of reliable reference genes for gene expression under abiotic stress in cotton (Gossypium hirsutum L.)

Reference genes are critical for normalization of the gene expression level of target genes. The widely used housekeeping genes may change their expression levels at different tissue under different treatment or stress conditions. Therefore, systematical evaluation on the housekeeping genes is required for gene expression analysis. Up to date, no work was performed to evaluate the housekeeping genes in cotton under stress treatment. In this study, we chose 10 housekeeping genes to systematically assess their expression levels at two different tissues (leaves and roots) under two different abiotic stresses (salt and drought) with three different concentrations. Our results show that there is no best reference gene for all tissues at all stress conditions. The reliable reference gene should be selected based on a specific condition. For example, under salt stress, UBQ7, GAPDH and EF1A8 are better reference genes in leaves; TUA10, UBQ7, CYP1, GAPDH and EF1A8 were better in roots. Under drought stress, UBQ7, EF1A8, TUA10, and GAPDH showed less variety of expression level in leaves and roots. Thus, it is better to identify reliable reference genes first before performing any gene expression analysis. However, using a combination of housekeeping genes as reference gene may provide a new strategy for normalization of gene expression. In this study, we found that combination of four housekeeping genes worked well as reference genes under all the stress conditions.