Nostoc, Microcoleus and Leptolyngbya inoculums are detrimental to the growth of wheat (Triticum aestivum L.) under salt stress

Background and aims

This study investigated the effect of cyanobacterial inoculants on salt tolerance in wheat.

Methods

Unicyanobacterial crusts of Nostoc, Leptolyngbya and Microcoleus were established in sand pots. Salt stress was targeted at 6 and 13 dS m^?1, corresponding to the wheat salt tolerance and 50 % yield reduction thresholds, respectively. Germinated wheat seeds were planted and grown for 14 (0 and 6 dS m^?1) and 21 (13 dS m^?1) days by which time seedlings had five emergent leaves. The effects of cyanobacterial inoculation and salinity on wheat growth were quantified using chlorophyll fluorescence, inductively coupled plasma-optical emission spectrometry and biomass measurements.

Results

Chlorophyll fluorescence was negatively affected by soil salinity and no change was observed in inoculated wheat. Effective photochemical efficiency correlated with a large range of plant nutrient concentrations primarily in plant roots. Inoculation negatively affected wheat biomass and nutrient concentrations at all salinities, though the effects were fewer as salinity increased.

Conclusions

The most likely explanation of these results is the sorption of nutrients to cyanobacterial extracellular polymeric substances, making them unavailable for plant uptake. These results suggest that cyanobacterial inoculation may not be appropriate for establishing wheat in saline soils but that cyanobacteria could be very useful for stabilising soils.     

Molecular (PCR-DGGE) versus morphological approach: analysis of taxonomic composition of potentially toxic cyanobacteria in freshwater lakes

Background

The microscopic Utermöhl method is commonly used for the recognition of the presence and taxonomic composition of potentially toxic cyanobacteria and is especially useful for monitoring reservoirs used as drinking water, recreation and fishery resources. However, this method is time-consuming and does not allow potentially toxic and nontoxic cyanobacterial strains to be distinguished. We have developed a method based on denaturing gradient gel electrophoresis (DGGE) of the marker gene ITS and the mcy-gene cluster, and DNA sequencing. We have attempted to calibrate the DGGE-method with a microscopic procedure, using water samples taken in 2011 from four lakes of the Great Mazurian Lakes system.

Results

Results showed that the classic microscopic method was much more precise and allowed the classification of the majority of cyanobacterial taxa to the species or genus. Using the molecular approach, most of the sequences could only be assigned to a genus or family. The results of DGGE and microscopic analyses overlapped in the detection of the filamentous cyanobacteria. For coccoid cyanobacteria, we only found two taxa using the molecular method, which represented 17% of the total taxa identified using microscopic observations. The DGGE method allowed the identification of two genera of cyanobacteria (Planktothrix and Microcystis) in the studied samples, which have the potential ability to produce toxins from the microcystins group.

Conclusions

The results confirmed that the molecular approach is useful for the rapid detection and taxonomic distinction of potentially toxic cyanobacteria in lake-water samples, also in very diverse cyanobacterial communities. Such rapid detection is unattainable by other methods. However, with still limited nucleotide sequences deposited in the public databases, this method is currently not sufficient to evaluate the entire taxonomic composition of cyanobacteria in lakes.     

Distribution of picophytoplankton communities from brackish to hypersaline waters in a South Australian coastal lagoon

Background

Picophytoplankton (i.e. cyanobacteria and pico-eukaryotes) are abundant and ecologically critical components of the autotrophic communities in the pelagic realm. These micro-organisms colonized a variety of extreme environments including high salinity waters. However, the distribution of these organisms along strong salinity gradient has barely been investigated. The abundance and community structure of cyanobacteria and pico-eukaryotes were investigated along a natural continuous salinity gradient (1.8% to 15.5%) using flow cytometry.

Results

Highest picophytoplankton abundances were recorded under salinity conditions ranging between 8.0% and 11.0% (1.3 × 10⁶ to 1.4 × 10⁶ cells ml^-1). Two populations of picocyanobacteria (likely Synechococcus and Prochlorococcus) and 5 distinct populations of pico-eukaryotes were identified along the salinity gradient. The picophytoplankton cytometric-richness decreased with salinity and the most cytometrically diversified community (4 to 7 populations) was observed in the brackish-marine part of the lagoon (i.e. salinity below 3.5%). One population of pico-eukaryote dominated the community throughout the salinity gradient and was responsible for the bloom observed between 8.0% and 11.0%. Finally only this halotolerant population and Prochlorococcus-like picocyanobacteria were identified in hypersaline waters (i.e. above 14.0%). Salinity was identified as the main factor structuring the distribution of picophytoplankton along the lagoon. However, nutritive conditions, viral lysis and microzooplankton grazing are also suggested as potentially important players in controlling the abundance and diversity of picophytoplankton along the lagoon.

Conclusions

The complex patterns described here represent the first observation of picophytoplankton dynamics along a continuous gradient where salinity increases from 1.8% to 15.5%. This result provides new insight into the distribution of pico-autotrophic organisms along strong salinity gradients and allows for a better understanding of the overall pelagic functioning in saline systems which is critical for the management of these precious and climatically-stress ecosystems.     

Isolation of ACC deaminase producing PGPR from rice rhizosphere and evaluating their plant growth promoting activity under salt stress

Aims

Bacteria possessing ACC deaminase activity reduce the level of stress ethylene conferring resistance and stimulating growth of plants under various biotic and abiotic stresses. The present study aims at isolating efficient ACC deaminase producing PGPR strains from the rhizosphere of rice plants grown in coastal saline soils and quantifying the effect of potent PGPR isolates on rice seed germination and seedling growth under salinity stress and ethylene production from rice seedlings inoculated with ACC deaminase containing PGPR.

Methods

Soils from root region of rice growing in coastal soils of varying salinity were used for isolating ACC deaminase producing bacteria and three bacterial isolates were identified following polyphasic taxonomy. Seed germination, root growth and stress ethylene production in rice seedlings following inoculation with selected PGPR under salt stress were quantified.

Results

Inoculation with selected PGPR isolates had considerable positive impacts on different growth parameters of rice including germination percentage, shoot and root growth and chlorophyll content as compared to uninoculated control. Inoculation with the ACC deaminase producing strains reduced ethylene production under salinity stress.

Conclusions

This study demonstrates the effectiveness of rhizobacteria containing ACC deaminase for enhancing salt tolerance and consequently improving the growth of rice plants under salt-stress conditions.     

滨海盐渍化土壤中蓝细菌多样性及分布

【目的】蓝细菌在贫瘠土壤的固氮、固碳中发挥着重要作用,然而土壤盐渍化对蓝细菌多样性及群落结构的影响还不清楚。本研究以莱州湾南岸及黄河口海水入侵盐渍化土壤为例,研究蓝细菌的多样性、群落结构及丰度沿盐度梯度的分布情况。【方法】利用自动核糖体间隔基因分析(ARISA)技术与群落相似性分析(ANOSIM)探究蓝细菌群落的差异与空间分布格局;通过16S rRNA基因克隆文库、测序与系统进化分析解析了3个典型盐度梯度样品中蓝细菌的群落组成;实时定量PCR测定蓝细菌16S rRNA拷贝数(丰度);BEST多元分析探寻影响蓝细菌分布的主要环境因子。【结果】蓝细菌在莱州湾南岸盐土中分布广泛,其群落结构在低(0.63%?1.27%)、中(1.55%?2.00%)、高盐度(2.39%?5.11%)样品组之间差异显著(P=0.03),而在不同含水量样品组间则不显著(P=0.09)。总体来看,群落结构主要受土壤盐度与含水量这两个因子联合控制(P=0.02)。低盐样品中蓝细菌多样性和丰富度最低,并以Halomicronema和Acaryochloris为优势类群。Leptolyngbya在中、高盐土中占优势。Arthrospira与Geitlerinema仅在低盐土中检测到,而Oscillatoria则仅出现在高盐土中。随盐度升高蓝细菌丰度呈下降趋势,低盐土样中蓝细菌丰度(2.14×105 copies/g干土)显著高于中盐土(1.25×105 copies/g干土)(P<0.05);高盐度土为1.20×105 copies/g干土。【结论】盐渍化程度是调控莱州湾南岸滨海土壤中蓝细菌群落结构与丰度的最重要环境因子,可能对滨海土壤微生物碳氮循环产生重要影响。     

Cyanobacterial Diversity in the Alkaline Lake Khilganta during the Dry and Wet Periods

Clone libraries and morphological analysis were used to investigate cyanobacterial diversity in the cyanobacterial mat and dry crust at the bottom of the shallow, saline, alkaline Lake Khilganta (Southern Siberia, Russia). Filamentous cyanobacteria belonging to Phormidium genus and Coleofasciculus chthonoplastes were found to predominate during the dry period (2006) and the wet periods (1995 and 2012), respectively. Community composition during the dry and wet periods differed significantly. While 11 operational taxonomic units of cyanobacteria were revealed, only 3 occurred during both dry and wet periods. Occurrence of cosmopolitan C. chthonoplastes, which is common in neutral saline environments, is not typical of a continental alkaline lake and may be explained by the similarity of the dominant ions composition in lake water and in seawater.     

Effect of selenium fertilization on the accumulation of cadmium and lead in rice plants

Background and Aims

The accumulation of cadmium and lead in rice (Oryza sativa L.) grains is a potential threat to human health. In this study, the effect of selenium fertilization on the uptake and translocation of cadmium and lead in rice plants was investigated.

Methods

Rice plants were cultivated using cadmium and lead contaminated soils with selenium addition at three concentrations (0, 0.5 and 1 mg kg^?1). At maturity, plants were harvested, and element concentrations in rice tissues were analyzed by using ICP-MS.

Results

Selenium application significantly increased selenium accumulation in rice grain, and markedly decreased cadmium and lead concentrations in rice tissues. In brown rice grains, selenium application reduced cadmium concentrations by 44.4 %, but had no significant effect on lead accumulation. Selenium application significantly decreased metal mobility in soils, at 0.5 mg kg^?1 treatment, the translocation factor of cadmium and lead from soil to iron plaque decreased by 71 and 33 % respectively.

Conclusions

The mechanism of selenium mitigating of heavy metal accumulation in rice could be decreasing metal bioavailability in soil. Selenium fertilization could be an effective and feasible method to enrich selenium and reduce cadmium levels in brown rice.     

Assessment of the relationship between geologic origin of soil, rhizobacterial community composition and soil receptivity to tobacco black root rot in Savoie region (France)

Background and aims

In Morens (Switzerland), soils formed on morainic deposits (which contain vermiculite clay and display particular tobacco rhizobacterial community) are naturally suppressive to Thielaviopsis basicola-mediated tobacco black root rot, but this paradigm was never assessed elsewhere. Here, we tested the relation between geology and disease suppressiveness in neighboring Savoie (France).

Methods

Two morainic and two sandstone soils from Savoie were compared based on disease receptivity (T. basicola inoculation tests on tobacco), clay mineralogy (X-ray diffraction), tobacco rhizobacterial community composition (16S rRNA gene-based taxonomic microarray) and phlD ⁺ Pseudomonas populations involved in 2,4-diacetylphloroglucinol production (real-time PCR and tRFLP).

Results

Unlike in Morens, in Savoie the morainic soils were receptive to disease whereas T. basicola inoculation did not increase disease level in the sandstone soils. Vermiculite was not present in Savoie soils. The difference in rhizobacterial community composition between Savoie morainic and sandstone soils was significant but modest, and there was little agreement in bacterial taxa discriminating soils of different disease receptivity levels when comparing Morens versus Savoie soils. Finally, phlD ⁺ rhizosphere pseudomonads were present at levels comparable to those in Morens soils, but with different diversity patterns.

Conclusions

The morainic model of black root rot suppressiveness might be restricted to the particular type of moraine occurring in the Morens region, and the low disease receptivity of sandstone soils in neighboring Savoie might be related to other plant-protection mechanisms.     

Possible amelioration of coastal soil salinity using halotolerant nitrogen-fixing cyanobacteria

A brackish-water, nitrogen-fixing cyanobacterium, Anabaena torulosa, could successfully grow and fix nitrogen on moderately saline Kharland soils (soil conductivity 5 to 8.50 dS m-1), typical of Indian coastline. During five weeks of growth under laboratory as well as field conditions, the cyanobacterium exhibited high rates of nitrogen fixation and substantially enriched the nitrogen status of saline soils (43-76%), although the fixed nitrogen remained confined to the cyanobacterial biomass. Most (>90%) of the cell-bound Na+ remained extracellularly trapped in the mucopolysaccharide sheath of A. torulosa; traces of the cation that permeated cyanobacterial cells were found to exist in an osmotically active, free state. No evidence was found for the incorporation of Na+ into any biomolecule, especialty proteins or carbohydrates. Therefore, permanent removal of Na+ from saline soils using cyanobacteria may not be possible, since Na+ is released back into the soil subsequent to the death and decay of cyanobacteria. Removal of top soil containing cyanobacterial mats significantly decreased the soil salinity (between 26-38%). But such a practice removes all the fixed nitrogen and carbon and also does not seem feasible on a large scale. Amelioration of soil salinity by simultaneous application of A. torulosa during crop growth seems to be an attractive possibility, especially since it can also supplement the nitrogen requirement of the crop.     

Cyanolichens can have both cyanobacteria and green algae in a common layer as major contributors to photosynthesis

Background and Aims

Cyanolichens are usually stated to be bipartite (mycobiont plus cyanobacterial photobiont). Analyses revealed green algal carbohydrates in supposedly cyanobacterial lichens (in the genera Pseudocyphellaria, Sticta and Peltigera). Investigations were carried out to determine if both cyanobacteria and green algae were present in these lichens and, if so, what were their roles.

Methods

The types of photobiont present were determined by light and fluorescence microscopy. Small carbohydrates were analysed to detect the presence of green algal metabolites. Thalli were treated with selected strengths of Zn²⁺ solutions that stop cyanobacterial but not green algal photosynthesis. CO₂ exchange was measured before and after treatment to determine the contribution of each photobiont to total thallus photosynthesis. Heterocyst frequencies were determined to clarify whether the cyanobacteria were modified for increased nitrogen fixation (high heterocyst frequencies) or were normal, vegetative cells.

Key Results

Several cyanobacterial lichens had green algae present in the photosynthetic layer of the thallus. The presence of the green algal transfer carbohydrate (ribitol) and the incomplete inhibition of thallus photosynthesis upon treatment with Zn²⁺ solutions showed that both photobionts contributed to the photosynthesis of the lichen thallus. Low heterocyst frequencies showed that, despite the presence of adjacent green algae, the cyanobacteria were not altered to increase nitrogen fixation.

Conclusions

These cyanobacterial lichens are a tripartite lichen symbiont combination in which the mycobiont has two primarily photosynthetic photobionts, ‘co-primary photobionts’, a cyanobacterium (dominant) and a green alga. This demonstrates high flexibility in photobiont choice by the mycobiont in the Peltigerales. Overall thallus appearance does not change whether one or two photobionts are present in the cyanobacterial thallus. This suggests that, if there is a photobiont effect on thallus structure, it is not specific to one or the other photobiont.     

Cyanobacterial diversity shifts induced by butachlor in selected Indian rice fields in Eastern Uttar Pradesh and Western Bihar analyzed with PCR and DGGE

The present study examines the effects of 30 mg/kg butachlor on the cyanobacterial diversity of rice fields in Eastern Uttar Pradesh and Western Bihar in India. A total of 40 samples were grouped into three classes [(i) acidic, (ii) neutral, and (iii) alkaline soils], based on physicochemical and principle component analyses. Acidic soils mainly harbored Westillopsis, Trichormus, Anabaenopsis, and unicellular cyanobacteria; whereas Nostoc, Anabaena, Calothrix, Tolypothrix, and Aulosira were found in neutral and alkaline soils. Molecular characterization using 16S rRNA PCR and DGGE revealed the presence of 13 different phylotypes of cyanobacteria in these samples. Butachlor treatment of the soil samples led to the disappearance of 5 and the emergence of 2 additional phylotypes. A total of 40 DGGE bands showed significant reproducible changes upon treatment with butachlor. Phylogenetic analyses divided the phylotypes into five major clusters exhibiting interesting links with soil pH. Aulosira, Anabaena, Trichormus, and Anabaenopsis were sensitive to butachlor treatment, whereas uncultured cyanobacteria, a chroococcalean member, Westillopsis, Nostoc, Calothrix, Tolypothrix, Rivularia, Gloeotrichia, Fischerella, Leptolyngbya, and Cylindrospermum, appeared to be tolerant against butachlor at their native soil pH. Butachlor-induced inhibition of nitrogen fixation was found to be 65% (maximum) and 33% (minimum) in the soil samples of pH 9.23 and 5.20, respectively. In conclusion, low butachlor doses may prove beneficial in paddy fields having a neutral to alkaline soil pH.     

Salt accumulation and depletion in the root-zone of the halophyte Atriplex nummularia Lindl.: influence of salinity,leaf area and plant water use

Background and aims

Salt is known to accumulate in the root-zone of Na⁺ excluding glycophytes under saline conditions. We examined the effect of soil salinity on Na⁺ and Cl^? depletion or accumulation in the root-zone of the halophyte (Atriplex nummularia Lindl).

Methods

A pot experiment was conducted in soil to examine Na⁺ and Cl^? concentrations adjacent to roots at four initial NaCl treatments (20, 50, 200 or 400 mM NaCl in the soil solution). Plant water use was manipulated by leaving plants with all leaves intact, removing approximately 50 % of leaves, or removing all leaves. Daily evapotranspiration was replaced by watering undrained pots to weight with deionised water. After 35-38 days, samples were taken of the bulk soil and of soil loosely- and closely-adhering to the roots.

Results

In plants with leaves intact grown with 200 and 400 mM NaCl, average Na⁺ and Cl^? concentrations in the closely adhering soil were about twice the concentrations of the bulk soil. Ion accumulation increased with final leaf area and with cumulative transpiration over the duration of the trial. By contrast, in plants grown with the lowest salinity treatment (20 mM NaCl), Na⁺ and Cl^? concentrations decreased in the closely adhering soil with increasing leaf area and increasing cumulative water use.

Conclusions

Our data show that Na⁺ and Cl^? are depleted from the root-zone of A. nummularia at low salinity but accumulate in the root-zone at moderate to high salinity, and that the ions are drawn towards the plant in the transpiration stream.     

Drying and wetting in saline and saline-sodic soils—effects on microbial activity, biomass and dissolved organic carbon

Aims

There are few studies on the interactive effect of salinity and sodicity in soils exposed to drying and wetting cycles. We conducted a study to assess the impact of multiple drying and wetting on microbial respiration, dissolved organic carbon and microbial biomass in saline and saline-sodic soils.

Methods

Different levels of salinity (EC_1:5 1.0 or 2.5) and sodicity (SAR?<?3 or 20) were induced by adding NaCl and CaCl₂ to a non-saline/non-sodic soil. Finely ground wheat straw residue was added at 20?g?kg^?1 as substrate to stimulate microbial activity. The constant moist (CM) treatment was kept at optimum moisture content for the length of the experiment. The drying and rewetting (DW) treatments consisted of 1 to 3 DW cycles; each DW cycle consisted of 1?week drying after which they were rewet to optimum moisture and then maintained moist for 1?week.

Results

Drying reduced respiration more strongly at EC2.5 than with EC1.0. Rewetting of dry soils produced a flush in respiration which was greatest in the soils without salt addition and smallest at high salinity (EC2.5) suggesting better substrate utilisation by microbes in soils without added salts. After three DW events, cumulative respiration was significantly increased by DW compared to CM, being 24% higher at EC1.0 and 16% higher at EC2.5 indicating that high respiration rates after rewetting may compensate for the low respiration rates during the dry phase. The respiration rate per unit MBC was lower at EC2.5 than at EC1.0. Further, the size of the flush in respiration upon rewetting decreased with each ensuing DW cycle being 50–70% lower in the third DW cycle than the first.

Conclusions

Both salinity and sodicity alter the effect of drying and rewetting on soil carbon dynamics compared to non-saline soils.     

Growth,physiological, biochemical and ionic responses of pistachio seedlings to mild and high salinity

Key message

Depending on salt concentrations, different mechanisms are involved in the tolerance of pistachio and an acclimation to salinity conditions occurs in the leaves that develop in the presence of salt.

Abstract

Pistachio (Pistacia vera L.) is a salt tolerant species that is considered an alternative crop for cultivation in salinzied orchard soils. In this work, 12-week-old pistachio seedlings cultivated in soil under greenhouse conditions were treated with five levels of salinity including control (0.63 dSm^?1), low (2 and 4 dSm^?1) and high (8 and 10 dSm^?1) salt concentrations for further 12 weeks. Plant growth parameters were not affected by mild salinity; a significant reduction was only observed from 8 dSm^?1. Considerable differences were observed between the young and mature leaves regarding osmotic and ionic stress effects of salt. Main compatible solutes were proline in mature leaves, proline and soluble sugars in young leaves, and soluble sugars and amino acids, other than proline, in roots. Concentration and content of Na in the leaves were not significantly increased at low levels of salinity and the K:Na and Ca:Na ratio of leaves were affected only by higher salt concentrations. Using the sequential extraction procedure for cell wall isolation, we observed that both absolute and relative amounts of Na in the cell wall fraction increased under low salinity, while decreased under higher levels of salt supply. Stable water relations, photochemistry and CO₂ assimilation rates particularly of young leaves, as well as ion homeostasis were mechanisms for maintenance of plants growth under mild salinity. Under severe saline conditions, the impaired ability of mature leaves for synthesis of assimilates, preferent allocation of carbohydrates to roots for maintenance of osmotic homeostasis and finally, reduction of protein synthesis caused growth inhibition in pistachio.     

High-Resolution Differentiation of Cyanobacteria by Using rRNA-Internal Transcribed Spacer Denaturing Gradient Gel Electrophoresis

For many ecological studies of cyanobacteria, it is essential that closely related species or strains can be discriminated. Since this is often not possible by using morphological features, cyanobacteria are frequently studied by using DNA-based methods. A powerful method for analysis of the diversity and dynamics of microbial populations and for checking the purity and affiliation of cultivated strains is denaturing gradient gel electrophoresis (DGGE). We realized high-resolution discrimination of a variety of cyanobacteria by means of DGGE analysis of sections of the internal transcribed spacer between the 16S and 23S rRNA genes (rRNA-ITS). A forward primer specific for cyanobacteria, targeted at the 3′ end of the 16S rRNA gene, was designed. The combination of this primer and three different reverse primers targeted to the rRNA-ITS or to the 23S rRNA gene yielded PCR products of different sizes from cultures of all 16 cyanobacterial genera that were tested but not from other bacteria. DGGE profiles produced from the shortest section of rRNA-ITS consisted of one band for all but one cyanobacterial genera, and those generated from longer stretches of rRNA-ITS yielded DGGE profiles containing one to four bands. The suitability of DGGE for detecting intrageneric and intraspecific variation was tested by using strains of the genus Microcystis. Many strains could be discriminated by means of rRNA-ITS DGGE, and the resolution of this method was strikingly higher than that obtained with previously described methods. The applicability of the developed DGGE assays for analysis of cyanobacteria in field samples was demonstrated by using samples from freshwater lakes. The advantages and disadvantages associated with the use of each developed primer set are discussed.     

Cyanobacteria inhabiting biological soil crusts of a polar desert: Sør Rondane Mountains,Antarctica

Molecular and morphological methods were applied to study cyanobacterial community composition in biological soil crusts (BSCs) from four areas (two nunataks and two ridges) in the Sør Rondane Mountains, Antarctica. The sampling sites serve as control areas for open top chambers (OTCs) that were put in place in 2010 at the time of sample collection and will be compared with BSC samples taken from the OTCs in the future. Cyanobacterial cell biovolume was estimated using epifluorescence microscopy, which revealed the dominance of filamentous cyanobacteria in all studied sites except the Utsteinen ridge, where unicellular cyanobacteria were the most abundant. Cyanobacterial diversity was studied by a combination of molecular fingerprinting methods based on the 16S rRNA gene (denaturing gradient gel electrophoresis (DGGE) and 454 pyrosequencing) using cyanobacteria-specific primers. The number of DGGE sequences obtained per site was variable and, therefore, a high-throughput method was subsequently employed to improve the diversity coverage. Consistent with previous surveys in Antarctica, both methods showed that filamentous cyanobacteria, such as Leptolyngbya sp., Phormidium sp. and Microcoleus sp., were dominant in the studied sites.In addition, the studied localities differed in substrate type, climatic conditions and soil parameters, which probably resulted in differences in cyanobacterial community composition. Furthermore, the BSC growing on gneiss pebbles had lower cyanobacterial abundances than BSCs associated with granitic substrates.     

Epilithic Cyanobacterial Communities of a Marine Tropical Beach Rock (Heron Island, Great Barrier Reef): Diversity and Diazotrophy

The diversity and nitrogenase activity of epilithic marine microbes in a Holocene beach rock (Heron Island, Great Barrier Reef, Australia) with a proposed biological calcification “microbialite” origin were examined. Partial 16S rRNA gene sequences from the dominant mat (a coherent and layered pink-pigmented community spread over the beach rock) and biofilms (nonstratified, differently pigmented microbial communities of small shallow depressions) were retrieved using denaturing gradient gel electrophoresis (DGGE), and a clone library was retrieved from the dominant mat. The 16S rRNA gene sequences and morphological analyses revealed heterogeneity in the cyanobacterial distribution patterns. The nonheterocystous filamentous genus Blennothrix sp., phylogenetically related to Lyngbya, dominated the mat together with unidentified nonheterocystous filaments of members of the Pseudanabaenaceae and the unicellular genus Chroococcidiopsis. The dominance and three-dimensional intertwined distribution of these organisms were confirmed by nonintrusive scanning microscopy. In contrast, the less pronounced biofilms were dominated by the heterocystous cyanobacterial genus Calothrix, two unicellular Entophysalis morphotypes, Lyngbya spp., and members of the Pseudanabaenaceae family. Cytophaga-Flavobacterium-Bacteroides and Alphaproteobacteria phylotypes were also retrieved from the beach rock. The microbial diversity of the dominant mat was accompanied by high nocturnal nitrogenase activities (as determined by in situ acetylene reduction assays). A new DGGE nifH gene optimization approach for cyanobacterial nitrogen fixers showed that the sequences retrieved from the dominant mat were related to nonheterocystous uncultured cyanobacterial phylotypes, only distantly related to sequences of nitrogen-fixing cultured cyanobacteria. These data stress the occurrence and importance of nonheterocystous epilithic cyanobacteria, and it is hypothesized that such epilithic cyanobacteria are the principal nitrogen fixers of the Heron Island beach rock.     

Chronic mild salinity affects source leaves physiology and productivity parameters of rice plants (Oryza sativa L., cv. Taipei 309)

Aim

In rice, the top two leaves are the major carbohydrate source during grain filling. Physiological performance of these leaves under salinity may allow estimate stress-induced yield loss.

Methods

Greenhouse grown rice plants (cv. Taipei 309) were subjected to 10 and 20 mM NaCl stress levels from germination till maturity. Plant development was measured at the flowering stage and yield parameters were quantified after complete ripening of panicles.

Results

Gas exchange in the main source leaves were not significantly affected by any of the stress levels. However, growth parameters as well as total metabolizable carbohydrates content, chlorophyll content (CCI), maximal efficiency of PSII photochemistry in dark-adapted state (F _v/F _m) and lipid peroxidation were significantly affected. Rice yield, measured as total panicle production, declined to 78 and 21 % of controls in 10 and 20 mM NaCl stress, respectively. Stress-induced yield loss was positively related with the decline in CCI, F _v/F _m and K⁺/Na⁺ ratio as well as with the increase in lipid peroxidation and total soluble carbohydrate contents.

Conclusions

Though the stress levels used in this work are below what is considered the minimal critical threshold of toxicity for rice, they induce significant negative effects on plant development and yield, when present along the whole plant life cycle.