Primary production of aquatic macrophytes and their epiphytes in two shallow lakes (Peipsi and Võrtsjärv) in Estonia

In shallow lakes with large littoral zones, epiphytes and submerged macrophytes can make an important contribution to the total annual primary production. We investigated the primary production (PP) of phytoplankton, submerged macrophytes, and their epiphytes, from June to August 2005, in two large shallow lakes. The production of pelagic and littoral phytoplankton and of the dominant submerged macrophytes in the littoral zone (Potamogeton perfoliatus in Lake Peipsi and P. perfoliatus and Myriopyllum spicatum in Lake Võrtsjärv) and of their epiphytes was measured using a modified ¹⁴C method. The total PP of the submerged macrophyte area was similar in both lakes: 12.4 g C m^?2 day^?1 in Peipsi and 12.0 g C m^?2 day^?1 in Võrtsjärv. In Peipsi, 84.2% of this production was accounted for by macrophytes, while the shares of phytoplankton and epiphytes were low (15.6 and 0.16%, respectively). In Võrtsjärv, macrophytes contributed 58%, phytoplankton 41.9% and epiphytes 0.1% of the PP in the submerged macrophyte area. Epiphyte production in both lakes was very low in comparison with that of phytoplankton and macrophytes: 0.01, 5.04, and 6.97 g C m^?2 day^?1, respectively, in Võrtsjärv, and 0.02, 1.93, and 10.5 g C m^?2 day^?1, respectively, in Peipsi. The PP of the littoral area contributed 10% of the total summer PP of Lake Peipsi sensu stricto and 35.5% of the total summer PP of Lake Võrtsjärv.     

Effect of salt stress on cucumber: Na+–K+ ratio, osmolyte concentration, phenols and chlorophyll content

A pot experiment with 17 diverse genotypes of cucumber with four levels of salt stress viz., 0, 2, 4 and 6 dS m^?1 was carried out during 2006. ANOVA revealed significant differences amongst genotypes and genotype × salt stress interaction indicating the genetic variability and differential response of the genotypes to different salt stress levels. The salt stress adversely affected the biochemical parameters; effects were severe under 4 dS m^?1. No genotype could survive at 6 dS m^?1. Sodium content, Na⁺–K⁺ ratio, proline, reducing sugars, phenol and yield reduction (%) increased significantly as the salt stress increased. Potassium, chlorophyll, membrane stability index and fruit yield decreased significantly under salt stress in all genotypes. However, the genotypes CRC-8, CHC-2 and G-338 showed lower accumulation of sodium, lesser depletion of potassium, lower Na⁺–K⁺ ratio and higher accumulation of proline, reducing sugars, phenols, better membrane stability and lower yield reduction (%) under salt stress, while CH-20 and DC-1 were sensitive to salt stress. Thus, a combination of traits such as higher membrane stability, lower Na⁺–K⁺ ratio, higher osmotic concentration and selective uptake of useful ions and prevention of over accumulation of toxic ions contribute to salt stress tolerance in cucumber. These traits would be useful selection criteria during salt stress breeding in cucumber.     

Probing the effect of electric field in 9,10-dimethoxy-2,6-bis(2-p-tolylethynyl)anthracene molecular nanowire using quantum chemical and charge density analysis

The effect of electric field (EF) in a newly designed molecular nanowire 9,10-dimethoxy-2,6-bis(2-p-tolylethynyl)anthracene has been analysed theoretically from the structural and electronic charge transport properties using quantum chemical and charge density calculations. The applied EF (0–0.36 VÅ^? 1) alters the molecular conformation, charge density distribution, electrostatic properties and the electronic energy levels of the molecule. Furthermore, the applied EF decreases the highest occupied molecular orbital–lowest unoccupied molecular orbital gap significantly from 1.775 to 0.258 eV and it also induces polarisation in the molecule, which leads to increase the dipole moment of the molecule. The electrostatic potential for various levels of applied EF reveals the charge-accumulated regions of the molecule. The I–V characteristics of the molecule have been studied against various applied fields using Landauer formalism.     

Intermittent aeration affects the bioremediation potential of two red algae cultured in finfish effluent

The high cost of aeration needed to tumble culture macroalgae is a limiting factor for integration with land-based finfish culture. Toward reducing this electricity cost, we compared intermittent aeration (16 h on:8 h off) with continuous aeration (24 h on) on the productivity of two strains of Chondrus crispus (Basin Head and Charlesville) and Palmaria palmata from Atlantic Canada between May and June 2011. Algal fronds were cultured under a 16:8-h light/dark photoperiod in 50-L tanks supplied with finfish effluent (49 μmol L^?1 of ammonium and 11 μmol L^?1 of phosphate) at a mean water flow rate of 0.4 L min^?1. Nitrogen (N) influx was 1.8 gN m^?2 day^?1, and phosphorus (P) influx was 0.9 gP m^?2 day^?1, with uptake rates ranging from 0.02 to 2.4 gN m^?2 day^?1 and ?0.2 to 0.4 gP m^?2 day^?1. On average, the macroalgae culture system (algae and biofilms) removed 1.0 gN m^?2 day^?1 (51.9 %). The growth of macroalgae (pooled across treatment and strain) ranged from 0.5 to 1.6 % day^?1, which accounted for a yield of 2.2 to 5.4 g DW m^?2 day^?1. Switching off aeration at night improved the growth rate of Basin Head Chondrus by 146 % and had no effect on growth rate or nitrogen and carbon removal by P. palmata and Charlesville Chondrus. Growth and yield of Basin Head Chondrus under intermittent aeration were over two times greater than both Charlesville Chondrus treatments.     

The effects of external electric field: creating non-zero first hyperpolarizability for centrosymmetric benzene and strongly enhancing first hyperpolarizability for non-centrosymmetric edge-modified graphene ribbon H2N-(3,3)ZGNR-NO2

How to generate a non-zero first hyperpolarizability for a centrosymmetric molecule is a challenging question. In this paper, an external (pump) electric field is used to make a centrosymmetric benzene molecule generate a non-zero value of the electric field induced first hyperpolarizability (β ^F ). This comes from the centrosymmetry breaking of electron cloud. Two interesting rules are exhibited. (1) β ^F is anisotropic for different directional fields (F _i, i?=?X, Y, Z). (2) The field dependence of β ^F is a non-monotonic function, and an optimum external electric field causes the maximum value of β ^F . The largest first hyperpolarizability β ^F reaches the considerable level of 3.9?×?10⁵ a.u. under F _Y?=?330?×?10^?4 a.u. for benzene. The external electric field effects on non-centrosymmetric edge-modified graphene ribbon H₂N-(3,3)ZGNR-NO₂ was also studied in this work. The first hyperpolarizability reaches as much as 2.1?×?10⁷ a.u. under F _X?=?600?×?10^?4 a.u. for H₂N-(3,3)ZGNR-NO₂. We show that the external electric field can not only create a non-zero first hyperpolarizability for centrosymmetric molecule, but also remarkably enhance the first hyperpolarizability for a non-centrosymmetric molecule.     

Effect of tillage system on the root growth of spring wheat

Little research has examined the influence of tillage system on root growth in wheat grown on rainfed Vertisols. A 3-year field study (2003, 2004 and 2005) was carried out on a typical Vertisol (southern Spain), to determine the effects of tillage system on root growth in spring wheat (Triticum aestivum L) grown in continuous rotation with faba bean (Vicia faba L), within the framework of the long-term “Malagón” experiment started in 1986. Tillage treatments were no-tillage (NT) and conventional tillage (CT), and the experiment was designed as a randomized complete block with three replications. The following parameters were measured: above-ground biomass, grain yield, root length density (RLD), root biomass (RB) and root N content. In the topmost 10 cm of soil, higher values were found under CT than under NT for RLD in the rainiest year (20.2 km m^?3 vs. 9.6 km m^?3 respectively) and for RB (512 kg ha^?1 vs. 261 kg ha^?1 respectively) in all study years. In deeper layers, no difference was recorded between the two tillage systems. Greater wheat root development in the upper soil layer under CT may reflect the greater soil penetration resistance found in the topmost 10 cm under NT. Root separation using a sieve with a 0.5 mm mesh screen led to a marked underestimation of RLD and RB, with values up to three times higher when using a 0.2 mm mesh screen. Mean wheat root N content in the topmost 30 cm of soil accounted for over 80% of total root N content. The highest grain yield was observed under NT, since this system provided greater water storage in the soil profile in the mostly dry study years.     

Immobilized Growth of the Peridinin-Producing Marine Dinoflagellate Symbiodinium in a Simple Biofilm Photobioreactor

Products from phototrophic dinoflagellates such as toxins or pigments are potentially important for applications in the biomedical sciences, especially in drug development. However, the technical cultivation of these organisms is often problematic due to their sensitivity to hydrodynamic (shear) stress that is a characteristic of suspension-based closed photobioreactors (PBRs). It is thus often thought that most species of dinoflagellates are non-cultivable at a technical scale. Recent advances in the development of biofilm PBRs that rely on immobilization of microalgae may hold potential to circumvent this major technical problem in dinoflagellate cultivation. In the present study, the dinoflagellate Symbiodinium voratum was grown immobilized on a Twin-Layer PBR for isolation of the carotenoid peridinin, an anti-cancerogenic compound. Biomass productivities ranged from 1.0 to 11.0 g m^?2 day^?1 dry matter per vertical growth surface and a maximal biomass yield of 114.5 g m^?2, depending on light intensity, supplementary CO₂, and type of substrate (paper or polycarbonate membrane) used. Compared to a suspension culture, the performance of the Twin-Layer PBRs exhibited significantly higher growth rates and maximal biomass yield. In the Twin-Layer PBR a maximal peridinin productivity of 24 mg m^?2 day^?1 was determined at a light intensity of 74 μmol m^?2 s^?1, although the highest peridinin content per dry weight (1.7 % w/w) was attained at lower light intensities. The results demonstrate that a biofilm-based PBR that minimizes hydrodynamic shear forces is applicable to technical-scale cultivation of dinoflagellates and may foster biotechnological applications of these abundant marine protists.     

Differential growth and yield responses of salt-tolerant and susceptible rice cultivars to individual (Na<Superscript>+</Superscript> and Cl<Superscript>−</Superscript>) and additive stress effects of NaCl

Negative impacts exerted by sodium (Na⁺) and chloride (Cl^?) ions individually as well their possible additive effects (under NaCl) were evaluated on growth and yield reductions in rice, besides investigating whether salt-tolerant genotypes respond differentially than their sensitive counterparts. Though both Na⁺ and Cl^? ions get accumulated in plant tissues under NaCl stress, most research has historically been aimed to decipher harmful effects induced by Na⁺ ions. Accordingly, physiological and molecular mechanisms involved in Cl^? toxicity are not clearly understood in crop plants. To address these issues, 65-day-old plants of two rice cultivars, Panvel-3 (tolerant) and Sahyadri-3 (sensitive) were subjected to Cl^?, Na⁺ and NaCl (each with 100 mM concentration and electrical conductivity of ≈10 dS m^?1) stress using soil-based systems. Stress conditions were maintained till harvesting of mature (128-day-old) plants. All three treatments induced substantial antagonistic effects on growth, dry mass, yield components (number of grains per panicle, length, width, thickness and weight of grain, along with the percentage of grains filled) and overall crop yield, with greater impacts under NaCl than its constituent ions. Salinity treatments caused an imbalance in reducing sugars, protein, starch and proline contents, with the greatest magnitude under NaCl. A negative correlation between Cl^?/Na⁺ accumulation and crop yield was witnessed, with higher severity on the sensitive cultivar. The overall magnitude of toxicity was observed highest in NaCl followed by Na⁺ and Cl^?, respectively, suggesting additive effects of constituent ions under NaCl. Both cultivars responded similarly; however, the tolerant cultivar, unlike the sensitive one, kept Na⁺:K⁺ ratio <1.0 and accumulated proline in response to salinity treatments used in this study.     

Effect of Pulsed Electric Field Treatments on Permeabilization and Extraction of Pigments from Chlorella vulgaris

The effect of pulsed electric field (PEF) treatments of different intensities on the electroporation of the cytoplasmatic membrane of Chlorella vulgaris, and on the extraction of carotenoids and chlorophylls were investigated. Staining the cells with propidium iodide before and after the PEF treatment revealed the existence of reversible and irreversible electroporation. Application of PEF treatments in the range of 20–25 kV cm^?1 caused most of the population of C. vulgaris to be irreversibly electroporated even at short treatment times (5 pulses of 3 µs). However, at lower electric field strengths (10 kV cm^?1), cells that were reversibly electroporated were observed even after 50 pulses of 3 µs. The electroporation of C. vulgaris cells by PEF higher than 15 kV cm^?1 and duration is higher than 15 µs increased significantly the extraction yield of intracellular components of C. vulgaris. The application of a 20 kV cm^?1 for 75 μs increased the extraction yield just after the PEF treatment of the carotenoids, and chlorophylls a and b 0.5, 0.7, and 0.8 times, respectively. However, further increments in electric field strength and treatment time did not cause significant increments in the extraction yield. The extraction of carotenoids from PEF-treated C. vulgaris cells after 1 h of the application of the treatment significantly increased the extraction yield in comparison to the yield obtained from the cells extracted just after the PEF treatment. After PEF treatment at 20 kV cm^?1 for 75 µs, extraction yield for carotenoids, and chlorophylls a and b increased 1.2, 1.6, and 2.1 times, respectively. A high correlation was observed between irreversible electroporation and percentage of yield increase when the extraction was conducted after 1 h of the application of PEF treatment (R: 0.93), but not when the extraction was conducted just after PEF treatment (R: 0.67).     

Wastewater treatment and biofuel production through attached culture of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis> in a porous substratum biofilm reactor

The feasibility of attached culture Chlorella vulgaris in a porous substratum biofilm reactor (PSBR) for simultaneous wastewater treatment and biofuel production was investigated. The characteristics, including algal biofilm growth, lipid yield, nutrient removal, and energy efficiency of the outdoor cultures, were investigated under the influence of both inoculum densities and the percent submerged area. A maximum biofilm productivity of 57.87 g m^?2 d^?1 with 81.9 % adhesion was achieved under optimal conditions (inoculum density of 18 g m^?2 and the percent submerged area of 5.7 %). The lipid content and lipid yield were 38.56 % and 27.25 g m^?2 d^?1, respectively. Meanwhile, the algae removed 99.95 % ammonia, 96.05 % total nitrogen (TN), and 99.83 % total phosphorus (TP). Further, the energy life cycle for the PSBR was analyzed. The biomass productivity per unit irradiance was up to 4.6 g MJ^?1 (photosynthetic efficiency of 10.65 %). The PSBR was considered to be economically feasible due to the net energy ratio of 1.3 (>1).     

Identification of large variation in the photosynthetic induction response among 37 soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.] genotypes that is not correlated with steady-state photosynthetic capacity

Irradiance continuously fluctuates during the day in the field. The speed of the induction response of photosynthesis in high light affects the cumulative carbon gain of the plant and could impact growth and yield. The photosynthetic induction response and its relationship with the photosynthetic capacity under steady-state conditions (P _max) were evaluated in 37 diverse soybean [Glycine max (L.) Merr.] genotypes. The induction response of leaf photosynthesis showed large variation among the soybean genotypes. After 5 min illumination with strong light, genotype NAM23 had the highest leaf photosynthetic rate of 33.8 µmol CO₂ m^?2 s^?1, while genotype NAM12 showed the lowest rate at 4.7 µmol CO₂ m^?2 s^?1. Cumulative CO₂ fixation (CCF) during the first 5 min of high light exposure ranged from 5.5 mmol CO₂ m^?2 for NAM23 to 0.81 mmol CO₂ m^?2 for NAM12. The difference in the induction response among genotypes was consistent throughout the growth season. However, there was no significant correlation between CCF and P _max among genotypes suggesting that different mechanisms regulate P _max and the induction response. The observed variation in the induction response was mainly attributed to ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activation, but soybean lines differing in the induction response did not differ in the leaf content of Rubisco activase α- and β-proteins. Future studies will be focused on identifying molecular determinants of the photosynthetic induction response and determining whether this trait could be an important breeding target to achieve improved growth of soybeans in the field.     

Contributions of DOC from surface and groundflow into Lake Võrtsjärv (Estonia)

Hydrological changes have the greatest impact on shallow lakes where they alter the water volume and lake depth noticeably. Dissolved organic carbon (DOC), which is markedly affected by hydrological factors, has an important role in many biogeochemical processes. The DOC load supplied to Lake Võrtsjärv, the second largest lake in Estonia, was studied on the scale of the subcatchments discharging into the lake. Daily discharges and biweekly or monthly DOC concentrations were measured close to the river outlets over the years 1990–2002. The stream flow data were separated into groundflow and surface flow by applying local minimum and recursive digital filtering methods. Constituent load estimation software, LOADEST, was used to estimate DOC concentrations and load. LOADEST performed well for three of the four rivers. The total estimated DOC load to Võrtsjärv from all four main rivers varied from 1,320 to 4,934 t year^?1. The average annual load over the 13-year period was 3,265 t year^?1 or 1.18 g C m^?2 year^?1. Baseflow separation analysis indicated that the DOC load originating from groundflow contributed 79% and 69% of the total load according to the digital filter and local minimum methods, respectively. The results of our study demonstrate the utility of linking the rating-curve method and baseflow separation to assess the allochthonous DOC load to Võrtsjärv both currently and under changing climatic conditions.     

Nonlinear electric properties of DNA solutions.

DNA solutions are shown to present a nonlinear electric behavior. This property is measured through the third harmonic current intensity, which appears when the solution is placed in a sinusoidal electric field of moderately high strength (about 100 V·cm^?1). The influence of different parameters has been examined: fundamental frequency, field strength, concentration, molecular weight, and conformation. By progressive sonication, it is shown that the harmonic current is linearly proportional to the DNA molecular weight, but under an M_r of approximately 10⁶, the nonlinear electrical property decreases sharply and its spectrum shows a drastic change. It is thought that the harmonic current is not related to an orientational phenomenon; an explanation based on the electrical deformation of the molecule is suggested.     

Variability of bio-optical parameters in two North-European large lakes

The bio-optical properties of some North-European large lakes were examined during 1995–2005 using field data and laboratory measurements. The key variables were optically active substances (OAS: chlorophyll, total suspended matter and dissolved organic matter), Secchi depth, and the “spectrometric” and diffuse light attenuation coefficients. Our main study sites were Lake Peipsi and Lake Võrtsjärv in Estonia, both eutrophic with mean Secchi depth below 3 m. The measured water parameters were compared with those obtained from two clear-water Swedish lakes, Lake Vänern and Lake Vättern. This comparison describes the bio-optical differences of the water in eutrophic and oligotrophic lakes. The variability of water parameters in the turbid Estonian lakes was rather high, e.g. the chlorophyll content varied from 1.8 to 102 mg m^?3 and the diffuse light attenuation coefficient from 0.92 to 6.5 m^?1. The change in water properties depends on the season and the biological activity of phytoplankton. We found no apparent long-time trend in water properties. Regression analysis showed that in the turbid Estonian lakes the optical properties were well correlated with chlorophyll and suspended matter, but not with dissolved organic matter. The highest determination coefficients (between 0.73 and 0.89) were obtained when the optical parameters were correlated with all three OAS together (multiple regressions). Our results concerning the variability and interconnections among bio-optical parameters in two Estonian large lakes illustrate the effect of OAS and light field on the ecological conditions of lakes in general.     

Photosynthetic capacity of the capsule wall and its contribution to carbon fixation and seed yield in castor (<Emphasis Type="Italic">Ricinus communis</Emphasis> L.)

Defoliation occurs in castor due to several reasons, but the crop has propensity to compensate for the seed yield. Photosynthetic efficiency in terms of functional (gas exchange and chlorophyll fluorescence) and structural characteristics (photosynthetic pigment profiles and anatomical properties) of castor capsule walls under light- and dark-adapted conditions was compared with that of leaves. Capsule wall showed high intrinsic efficiency of photosystem II (F _v/F _m, 0.82) which was comparable to leaves (F _v/F _m, 0.80). With increasing photon flux densities (PFD), actual quantum yields and photochemical quenching coefficients of the capsule walls were similar to that in leaves, while electron transport rates reached a maximum corresponding to about 118 % of the leaves. However, maximum net photosynthetic rate of the capsule walls (2.60 µmol CO₂ m^?2 s^?1) was less than one-fourth of the leaves (15.67 µmol CO₂ m^?2 s^?1) at the CO₂ concentration of 400 µmol mol^?1, and the difference was attributed to about 80 % lower stomatal density and the 75 % lower total chlorophyll content of capsule walls than the leaves. Furthermore, seed weight in dark-adapted capsules was 2.70–12.42 % less as compared to the capsules developed under light. The results indicate that castor capsule walls are photosynthetically active (about 15–30 % of the leaves) and contribute significantly to carbon fixation and seed yield accounting for 10 % photoassimilates towards seed weight.     

Irradiance optimization of outdoor microalgal cultures using solar tracked photobioreactors

Photosynthetic activity and temperature regulation of microalgal cultures (Chlorella vulgaris and Scenedesmus obliquus) under different irradiances controlled by a solar tracker and different cell densities were studied in outdoor flat panel photobioreactors. An automated process control unit regulated light and temperature as well as pH value and nutrient concentration in the culture medium. CO₂ was supplied using flue gas from an attached combined block heat and power station. Photosynthetic activity was determined by pulse amplitude modulation fluorometry. Compared to the horizontal irradiance of 55 mol photons m^?2 d^?1 on a clear day, the solar tracked photobioreactors enabled a decrease and increase in the overall light absorption from 19 mol photons m^?2 d^?1 (by rotation out of direct irradiance) to 79 mol photons m^?2 d^?1 (following the position of the sun). At biomass concentrations below 1.1 g cell dry weight (CDW) L^?1, photoinhibition of about 35 % occurred at irradiances of ≥1,000 μmol photons m^?2 s^?1 photosynthetic active radiation (PAR). Using solar tracked photobioreactors, photoinhibition can be reduced and at optimum biomass concentration (≥2.3 g CDW L^?1), the culture was irradiated up to 2,000 μmol photons m^?2 s^?1 to overcome light limitation with biomass yields of 0.7 g CDW mol photons^?1 and high photosynthetic activities indicated by an effective quantum yield of 0.68 and a maximum quantum yield of 0.80 (F _v/F _m). Overheating due to high irradiance was avoided by turning the PBR out of the sun or using a cooling system, which maintained the temperature close to the species-specific temperature optima.     

2-Methylisoborneol production characteristics of <Emphasis Type="Italic">Pseudanabaena</Emphasis> sp. FACHB 1277 isolated from Xionghe Reservoir,China

The cyanobacterium Pseudanabaena sp. FACHB 1277, a 2-methylisoborneol (2-MIB) producer isolated from Xionghe Reservoir, was identified by molecular biological methods based on the 16S rDNA sequence. Pseudanabaena sp. FACHB 1277 is a planktonic freshwater species with relatively high 2-MIB per cell density value (7.76?×?10^?6 ng cell^?1) and specific growth rate (0.25?±?0.01 d^?1). The effects of temperature and light intensity on 2-MIB production of Pseudanabaena sp. FACHB 1277 were investigated. Of the six temperatures tested, 10, 15, 20, 25, 30, and 35 °C, the maximum total 2-MIB per cell density and minimum cell density were observed at 10 °C, while the total 2-MIB and dissolved 2-MIB (including extracellular and dissolved intracellular 2-MIB) increased with increasing temperature. Among the six tested light intensities (10, 25, 40, 55, 70, and 85 μmol photons m^?2 s^?1), the minimum total 2-MIB per cell density and maximum cell density were observed at 25 μmol photons m^?2 s^?1. The total 2-MIB and extracellular 2-MIB increased with light intensity increasing from 10 to 40 μmol photons m^?2 s^?1, while no significant increase was observed when the light intensity was higher than 40 μmol photons m^?2 s^?1. The maximum intracellular 2-MIB (including dissolved and bound) occurred at 25 μmol photons m^?2 s^?1. The present study indicates that increasing temperature could favor the conversion of bound intracellular to dissolved 2-MIB, while increasing light intensity stimulates the release of dissolved intracellular 2-MIB into the environment.     

Cas9-guide RNA ribonucleoprotein-induced genome editing in the industrial green alga <Emphasis Type="Italic">Coccomyxa</Emphasis> sp. strain KJ

Background

Oxygen-evolving photosynthetic microorganisms, collectively termed as microalgae, are gaining attention as alternative fuel sources. The unicellular alga Coccomyxa sp. strain KJ that belongs to the class Trebouxiophyceae can grow rapidly in minimal mineral media and accumulate triacylglycerols at levels?>?60% (w/w) of its dry weight under nitrogen depletion conditions. Thus, the strain can be a good candidate for biofuel production. Still, substantial improvements in lipid productivity and other traits of this strain are needed to meet commercial production requirements. Consequently, the development of new genetic tools including genome editing that are applicable to this strain is highly desired.

Results

In this paper, we report successful genome editing of strain KJ by intracellular delivery of a ribonucleoprotein complex comprising recombinant Cas9 protein and guide RNA. For introduction of Cas9-guide RNA ribonucleoprotein into strain KJ cells, we used an electroporator with a short (2.5 ms) electric pulse at a high field strength (7500 V cm^?1) followed by multiple 50-ms electric pulses at low field strength (250 V cm^?1). Under these conditions, we successfully isolated several knockout lines of the FTSY gene of strain KJ, encoding a signal recognition particle-docking protein at a frequency of 0.01%.

Conclusions

Our study shows applicability of DNA-free genome editing in Coccomyxa, which may be applicable in other Trebouxiophyceae species.

     

Salinity and drought interaction in wheat (Triticum aestivum L.) is affected by the genotype and plant growth stage

Salinity and drought are important agro-environmental problems occurring separately as well as together with the combined occurrence increasing with time due to climate change. Screening of bread wheat genotypes against salinity or drought alone is common; however, little information is available on the response of wheat genotypes to a combination of these stresses. This study investigates the response of a salt-resistant (SARC-1) and a salt-sensitive (7-Cerros) wheat genotype to drought at different growth stages under non-saline (ECe 2.1 dS m^?1) and saline soil (ECe 15 dS m^?1) conditions. Drought was applied by withholding water for 21 days at a particular growth stage viz. tillering, booting, and grain filling stages. At booting stage measurements regarding water relations, leaf ionic composition and photosynthetic attributes were made. At maturity grain yield and different yield, components were recorded. Salinity and drought significantly decreased grain yield and different yield components with a higher decrease in the case of combined stress of salinity × drought. The complete drought treatment (drought at tillering + booting + grain filling stages) was most harmful for wheat followed by drought at booting stage and grain filling–tillering stages, respectively. The salt-resistant wheat genotype SARC-1 performed better than the salt-sensitive genotype 7-Cerros in different stress treatments. A decrease in the water and turgor potentials, photosynthetic and transpiration rates, stomatal conductance, leaf K⁺, and increased leaf Na⁺ were the apparent causes of growth and yield reduction of bread wheat due to salinity, drought, and salinity × drought.