e a lower abiotic stress ( Bertness and Callaway, 1994) This dr

e. a lower abiotic stress ( Bertness and Callaway, 1994). This driver is expected to be different in TAE with a higher vegetation density consecutive to

the absence of durable snowbeds and Cabozantinib ic50 a continuous vegetative period. In two studies along TAE gradients, decreasing vegetation cover was correlated with amplified facilitative interactions among plants ( Smith, 1984 and Anthelme et al., 2012). While disentangling the respective influences on plant–plant interactions of all these abiotic parameters would require future studies, two hypotheses related to stress and disturbance sound particularly relevant to be tested in TAE. First, according to the SGH, increased aridity is expected to generate a higher frequency of facilitative interactions among plants, up to a threshold where Ixazomib chemical structure competitive effects predominate in the interactions (Michalet et al., 2006 and Maestre et al., 2009). Accordingly, a hypothesis to be tested is whether the stress–interaction

relationship is similar to that found along aridity gradients (with a reduction of facilitative interactions at the extremity of the gradient of stress) or whether the stress–interaction relationship rather resembles to that along altitudinal gradients in alpine environments, with an increase or at least a stabilization of the frequency of positive interactions with increasing stress (Choler et al., 2001 and le Roux and McGeoch, 2008). Second, variations Casein kinase 1 in frost-heaving regime is likely to alter the outcome of plant–plant interactions, with more facilitative effects observed either with stronger frost-heaving amplitudes (Venn et al., 2009) or with higher frost heaving frequency (Smith, 1981 and Pérez, 1987a). From this viewpoint, determining to what extent the amplitude of frost heaving (stronger in seasonal environments; Francou et al., 2001) or their frequency (higher in aseasonal environments) drive the outcome of plant–plant interactions

in TAE is a stimulating, unresolved challenge. Niche differentiation-related drivers of plant–plant interactions include at least four factors that are expected to vary between extratropical alpine environments and TAE (Fig. 1). First, facilitator’s size, which is expected to increase the frequency of facilitative interactions (Callaway and Walker, 1997), is larger in TAE than in extratropical alpine environments and associated with a higher ratio of aboveground to belowground biomass (Smith, 1994). A striking example of giant growth forms in TAE (Rundel, 1994) is giant rosettes such as Puya raimondii, which reach up to 4–6(12) m high ( Sgorbati et al., 2004). This “facilitator size” hypothesis seems to be corroborated by frequent observations of positive effects of giant rosettes on other plants ( Table 1: Smith, 1981, Smith, 1984, Pérez, 1987a, Young and Peacock, 1992 and Smith and Young, 1994).

The differences between the stations located close to populated a

The differences between the stations located close to populated areas were related mainly to the distribution of two families in a sample. Podoviridae Selleck AZD2281 (47.7%) and Myoviridae (37.9%) contributed mostly to the differences between groups 1 and 3, Siphoviridae (46.4%) and Podoviridae (43.3%) to the differences between groups 1 and 4, and Siphoviridae (46.2%) to the differences between groups 3 and 4. Significant

differences were observed between all the groups located close to populated areas and the groups in offshore stations in the lagoon (p < 0.05). In general, tailed phages made up more than 97% of the total number of phages detected, and long-tail phages were dominant, with tail lengths from 20 nm to 630 nm (Table 1). Phages with isometric heads were more frequent than prolate phages, and phages with contractile tails were more frequent than phages with non-contractile tails. In earlier reports all phages were considered to form size groups (Bratbak et al. 1990, Cochlan et al. 1993, Mathias et al. 1995, etc.). We placed all the observed phages into 5 size classes (30–60 nm; 60–80 nm; 80–100 nm; 100–120 nm; 120–160 nm), and the relative distribution of these classes was examined at all the study sites. Cluster analysis (75% Bray-Curtis similarity)

revealed that all the study sites in the Curonian Lagoon could be divided into three different groups corresponding to size classes (Figure 4) or three zones corresponding to geographical distribution. Group I, which was dominated selleck chemical Casein kinase 1 by the 30–60 nm and

60–80 nm size fractions, covered 4 stations with elevated water salinity recorded at the time of the study, which shows that mixing with different water bodies took place. Groups II and III represented the distribution of capsid sizes in the freshwater part of the lagoon. Group III covered two stations located in the open part of the lagoon and was dominated by the 30–60 nm size fraction (up to 48%). In group II, the 30–60 nm size fraction did not exceed 10%; the group was dominated by 80–100 nm and 100–120 nm capsid size phages. Both the latter size fractions constituted from 48% to 70% per station respectively. Phage-like particles of 200 nm capsid size (Figure 2aa) were found at stations 1, 8 and 11 with respective frequencies of 1, 1 and 2. These phages were not included in the cluster analysis as outliers. Analysis of size class contributions (SIMPER) to the differences between groups (in Figure 4) revealed that group I (sea water) differed from group II (freshwater) mainly in the 30–60 nm capsid size fraction (57.2%). Differences between the conditionally marine group I and the freshwater group III were due to 80–100 nm (34.9%) capsid phages. The difference between the two freshwater groups was due to the much higher relative abundance of 30–60 nm size fraction phages in group III (52%). Analysis of similarity (ANOSIM, based on Bray-Curtis similarity) revealed significant differences between groups I and III and between groups II and III (p < 0.

All primers for real-time RT-PCR are listed in Table S1 and Table

All primers for real-time RT-PCR are listed in Table S1 and Table S2. To determine the miRNA cleavage sites in the target genes, RLM-RACE was performed using the SMARTer RACE cDNA Amplification Kit (Clontech, PT4096-2). First, total RNA was extracted from the two tissues and ligated with SMARTer II A oligonucleotide, and then the RNA was reverse transcribed using 10 × Universal Primer A Mix (UPM). PCR was then performed twice, using the UPM/gene-specific primer in the first reaction and the UPM/nested gene-specific primer in the second, according to the manufacturer’s instructions. The product was then gel-extracted and cloned Selleck Docetaxel into

the PMD20-T Vector (Takara, Dalian, China) for sequencing. The primers for RLM-RACE are shown in Table S3. To investigate the small RNA expression profiles of O. longistaminata, two cDNA libraries of small RNAs, one from ASs and one from rhizomes, were sequenced. In total, 20,358,337 raw reads from ASs and 21,313,971 from rhizomes were produced. After 17-AAG nmr elimination of low quality reads,

adaptors and contaminating sequences, 17,547,018 and 18,655,858 clean reads with lengths of 17 to 30 nt remained from the ASs and rhizomes, respectively. Of these reads, 4,866,476 and 6,517,161, respectively, were unique ( Table 1). The overall size distributions of the sequenced reads from the two libraries were very similar, with the 24 nt class being the most abundant ( Fig. 1). The unique sequences were mapped to the rice genome assembly using Bowtie [22]. As shown in Table 1, almost every category of RNAs, including miRNAs, siRNAs, rRNAs, snoRNAs, snRNAs, tRNAs, repeat-associated sRNAs, and degraded mRNAs, were detected in both tissues. Finally, 11,265 and 33,536 reads for ASs, and 12,997 and 40,126 reads for rhizomes were identified as known and predicted miRNA candidates, respectively, for analysis. All small RNA sequences were searched against the plant miRNA database to identify known, conserved and novel miRNAs in ASs and rhizomes, as described in Materials and methods. To reduce false-positive rates, only sequences with at least two detected

reads were designated as miRNA candidates. Of the 713 known rice miRNAs deposited in the miRBase database (Release 20, June 2013), Dimethyl sulfoxide 380 known rice miRNAs were identified as being expressed in ASs and rhizomes, including 340 miRNAs found in both tissues (Table 2). Among them, 21 and 19 known miRNAs were expressed exclusively in ASs and in rhizomes, respectively (Fig. 2, Tables 2, S4). The most highly tissue-specific miRNAs included osa-miRNA319a-3p and osa-miRNA529a in the rhizomes and osa-miRNA530-5p and osa-miRNA5073 in the ASs, indicating their roles in rhizome and AS growth. In the conserved and novel miRNAs 72 conserved miRNAs were expressed, including 53 miRNAs common to ASs and rhizomes. Seven and 12 miRNAs were expressed specifically in ASs and rhizomes, respectively (Table S5).

The recipient was able to perceive phosphenes from stimulation of

The recipient was able to perceive phosphenes from stimulation of 39/80 electrodes, with the nature of elicited percepts varying from discrete, clustered, diffuse and elongated points this website of light. While the system was ultimately of no practical use to the recipient, it demonstrated that stimulation of visual cortex with a fully-implanted,

multi-electrode implant was feasible and could produce visuotopically organized percepts. Moreover, it suggested that with an increased number of electrodes and phosphenes, providing the blind with useful visual perception via cortical stimulation may be possible. Brindley׳s success was followed by a significant increase in research effort towards the selleck chemicals development of a cortical visual prosthesis, with a number of other groups publishing the results of experiments directed at visual prosthesis development in subsequent years (Dobelle and Mladejovsky, 1974, Pollen, 1975 and Talalla et al., 1974). Both Brindley and Dobelle׳s groups separately progressed their implants, eventually demonstrating that phosphenes could be assembled into simple patterns for the purpose of reading Braille characters (Brindley and

Rushton, 1974 and Dobelle, 1974). The goals of visual prosthesis designers at the time were generally centered on providing the blind with the ability to read text or to improve their level of independent mobility. Brindley had previously determined that 50 favorably placed phosphenes would be required to permit the reading of idealized letterforms, however up to 600 would be required for the reading of handwriting (Brindley, 1965). Dobelle was less focused on reading and

more so on mobility (Dobelle et al., 1979a) and his group reported plans to implant 512 electrodes towards that goal (Dobelle et al., 1979a). Brindley made no further reports on his visual prosthesis development program after 1982 (Brindley, 1982), however Dobelle continued Niclosamide development, incorporating a camera and miniaturizing the system to the point of portability. With ongoing improvements in the sophistication of computing hardware, Dobelle׳s (2000) system was ultimately capable of providing limited object recognition and mobility to one of its recipients with only 21 phosphenes. Despite this, it was similar to Brindley׳s device, based on an array of surface electrodes that required currents up to several milliamperes to elicit phosphenes (Brindley and Lewin, 1968). Aside from limiting the minimum spacing between electrodes and therefore any resultant phosphenes, these large currents also increased the risk of seizures for implant recipients (Naumann, 2012), a problem that had previously plagued other researchers in the field (Pudenz, 1993). A further disadvantage was the bulk of the cabling that connected to the electrodes via a transcutaneous connector fixed to the skull.

The biological method besides being more specific and efficient t

The biological method besides being more specific and efficient than thermal treatment can result in products of economical interest (e.g. enzymes, mushrooms, animal feed). Pleurotus ostreatus has been used in the bioremediation of pollutants and the degradation of lignocellulosic residue by the action of different enzymes ( Dundar, Acay, & Yildiz, 2009; Haritash & Kaushik, 2009), including the lignocellulolytic enzymes, tannase and phytase ( Batra & Saxena, 2005; Cavallazzi, Brito, Oliveira, Villas-Bôas, & Kasuya, 2004; Collopy & Royse, 2004).

In addition, this fungus produces mushrooms using different lignocellulosic residues ( Dundar et al., 2009; Fan, Soccol, Pandey, Vandenberghe, Bleomycin solubility dmso & Soccol, 2006; Nunes et al., 2012). The P. ostreatus mushrooms have high nutritional value and are sources of protein, carbohydrates, vitamins (e.g. B1, B2 and B3), calcium and iron ( Dundar et al., 2009; Wang, Sakoda, & Suzuki, 2001). Major agroindustrial residues have in its chemical composition higher fibers content with low availably than protein, minerals and vitamins (Villas-Bôas, Esposito, &

Mitchell, 2002). Colonization and solid fermentation Everolimus clinical trial by fungi have been used to increase the availably and the nutritional value of these residues (Pereira, 2011; Sánchez, 2009; Villas-Bôas et al., 2002). This procedure has been used with success in cocoa (Alemawor, Dzogbefia, Oddoye, & Oldham, 2009), sawdust (Kwak, Jung, & Kim, PI-1840 2008) and jatropha seed cake (Pereira, 2011). Thus, in this study, we tested the ability of P. ostreatus to degrade antinutritional

factors and produce edible mushrooms using different proportions of the J. curcas seed cake as substrate. The isolate Plo 6 of P. ostreatus, which were used in this study, belong to collection of the Department of Microbiology of Federal University of Viçosa, MG, Brazil. This isolate was grown in a Petri dish containing potato dextrose agar culture medium (Merck) at pH 5.8 and incubated at 25 °C. After 7 days, the mycelium was used for inoculum production (spawn) in a substrate made of rice grains with peel ( de Assunção et al., 2012). The rice grains were cooked for 30 min in water at a 1:3 (rice grains:water, w/w). After cooking, the grains were drained and supplemented with 0.35 (g/100 g) CaCO3 and 0.01 (g/100 g) CaSO4. These grains (70 g) were packed into small glass jars and sterilized in an autoclave at 121 °C for 1 h. After cooling, each jar was inoculated with 4 agar discs (5 mm diameter) containing mycelium and incubated in the dark at 25 ± 2 °C for 15 d. The J. curcas seed cake used in this study was obtained from an industry of biodiesel (Fuserman Biocombustíveis, Barbacena, Minas Gerais State, Brazil). The proper substrate composition for optimal growth and enzyme production by P. ostreatus was chosen based on previously experiments with jatropha seed cake and different agroindustrial residues ( Da Luz, 2009).

Phosphodiester models and mimics have been used widely to

Phosphodiester models and mimics have been used widely to selleck inhibitor understand the mechanisms

of phosphodiesterases such as nucleases and ribozymes. This section discusses examples where one or more of the bridging or non-bridging oxygen atoms associated with the phosphodiester group has been exchanged for either sulfur or fluorine. The resulting analogues are often reactive, where their altered reactivity profiles are used to probe the nature of catalysis in enzyme active sites and/or binding to metal ions therein. Some of the most poignant recent additions to the mechanistic toolbox are the phosphorothiolates (Table 2, entry 1), where a bridging oxygen atom has been replaced by sulfur. These systems have received significant attention because synthetic advances have permitted their use in oligonucleotides [14, 15 and 16]. Phosphorothiolates can also elucidate O-Mg2+ ion interactions through soft metal ion GSI-IX chemical structure rescue experiments. More significantly, where a leaving

group oxygen is replaced by sulfur, the enhanced leaving group properties of thiolate anions accelerate their departure, sometimes obviating the need for catalysis, and potentially making previously kinetically silent processes rate-determining. In this vein, phosphorothiolate studies have illuminated HDV [17•] and VS [18] ribozyme systems alongside nucleobase substitutions to provide unequivocal evidence in support of general acid/base catalysis. Recent work in this area, primarily from the Piccirilli laboratory, has been reviewed [19•• and 20]. More subtle substitution of phosphodiesters can be effected through the use of isotopomeric compounds, such as 18-O

labelled species (Table 2, entry 2). Heavy atom isotope effects are challenging to determine aminophylline on a practical level, however, isotopic substitutions represent the least perturbing of all possible analogues. 5′-18O and 2′-18O isotopomeric analogues of the dinucleotide 5′-UpG-3′ were synthesised and the base-promoted cleavage kinetics of these phosphodiester systems were explored [21••]. Through these studies, the transition state for the 2′-O-transphosphorylation process was suggested to be late in nature, and solvent deuterium isotope effect studies suggest the prior formation of the 2′-alkoxide nucleophile rather than rate-determining general base catalysis by hydroxide ion. An extension of this, supplemented with computational studies, has been used to revisit the mechanism of ribonuclease A [22]. Fluorophosphonates present the possibility of concerted, diester-like transition states while offering the size and hydrogen bonding characteristics of monoesters [23]. This mixed character was used to explore the promiscuous proficiencies of phosphoryl transfer by alkaline phosphatase.

Each individual quota is a portion of the total quota (Total Allo

Each individual quota is a portion of the total quota (Total Allowable Catches, TAC) that can be caught each year according to

the state of a stock. However, also in this case, the partners complained that small-scale fishermen are facing several difficulties in the access to these quotas: 90% of bluefin tuna national quota is hold by just a few big vessels, and the small-scale fisheries segment has access to just 10% of the authorized catches. Corsica Region adds that no fishing vessels in their fleet would be eligible for a TFC Selleckchem SGI-1776 system. In certain European areas (e.g. Scotland, Iceland) ITQs are mainly assigned on the basis of fishing vessels’ catch histories (species and quantities caught in recent years by each vessel); when it comes to new entries, quotas should be assigned taking into account the amounts that are allocated to vessels with similar characteristics. TACs are calculated for each target species on the basis of biological indices. Landings should be constantly monitored so that the fishing season can be terminated as soon as the TAC value is reached. Through this measure, FK866 molecular weight the management authority can fix lower catch levels if the resource is overexploited, so that stocks can progressively recover. Once a stock has reached sustainable levels again, TAC can be raised to the

initial or even higher values. However, two basic requirements must be satisfied for the

measure to be effective: on the one hand, catches should be constantly monitored (resource state assessment), on the other hand, fishermen should be constantly monitored too (compliance with NADPH-cytochrome-c2 reductase the rules). The TAC to be distributed among individual quota owners can only be determined if the state of stocks is known. None of the partners reckons that a system based on catch histories would be appropriate and feasible for the Mediterranean. The main reason is a general lack of sound and reliable individual historical data. The assessment of the status of resources is considered as a key factor for the introduction of management systems based on TFC. In particular TACs can only be determined on the basis of stock assessment data and models, but these are available only for a limited number of species in the Mediterranean Sea. In the Mediterranean Sea the use of TACs is no guarantee of success and of optimal management, since the two requirements mentioned above (resource assessment and compliance control) are not always completely satisfied. In the Mediterranean Sea a further criticality is related to the fact that demersal and pelagic stocks are shared among different States and this should be taken into account when assigning TFCs. The Adriatic Sea is probably the largest and best-defined area of occurrence of shared stocks in the Mediterranean.

Maureen will be remembered not only as an outstanding leader in t

Maureen will be remembered not only as an outstanding leader in the field of bone biology but also as a wonderful friend selleck and colleague to all who knew her. Maureen was born in the small village

of Benburb, County Tyrone, Northern Ireland as the middle child of five children of Robert and Elizabeth Howard who were teachers at the local village schools. Education was of paramount importance and she was awarded a scholarship to attend Methody School in Belfast. She excelled in Physics and was offered a bursary to study at Queen’s University, Belfast where she gained a first class B.Sc. Hons degree in Experimental Physics in 1948. Following a year at the Atomic Energy Research Establishment at Harwell, she moved to the University of Oxford, became a member of St Hugh’s College, met her

husband John and spent the rest of her career. She gained her DPhil degree in 1952 in Nuclear Physics at the Clarendon Laboratory, Oxford for research on “The Talazoparib mw investigation of nuclear reactions using photographic plate technique. Maureen’s earliest research papers dealt with Nuclear Physics and radiation measurement. Included was the determination of the efficiency of production of neutrons by deuterium–deuterium (D–D) interaction whilst studying for her D.Phil degree at the Clarendon laboratory, which was headed by Frederick Alexander Lindemann, Lord Cherwell, F.R.S. This background 3-oxoacyl-(acyl-carrier-protein) reductase in Nuclear Physics and investigations on the D–D reaction by using nuclear track emulsions to record the energy and angular distribution of 3Helium and 3H nuclei and thus neutron flux served her well in her later and prominent work on autoradiographic detection of radioisotopes in biological materials. After gaining

her higher degree, Maureen moved to the Nuffield Institute for Medical Research. This University of Oxford department had been established by Sir William Morris, later Lord Nuffield, in the Radcliffe Observatory, now part of Green Templeton College. Maureen always retained fond memories of her short time there in this exquisite central location in Oxford. The buildings had been purchased from the Radcliffe Trustees in 1934 following the erection of a new observatory in Pretoria, South Africa. It was an idyllic place to work as the old Observatory was a beautiful building that had been constructed in “a calm and retired locality” and has been described as the finest Georgian building in Oxford. Its construction as an observatory was completed in 1794 to the designs of James Wyatt, based on a small Tower of the Winds in Athens from the benefaction of Dr John Radcliffe. Geoffrey Dawes, a foetal physiologist, had become the director of the Nuffield Institute for Medical Research in Oxford in 1948 and provided a small room for use of microradiographic equipment by Dame Janet Vaughan’s staff; he and Maureen remained firm friends throughout her time in Oxford.

The extents of fresh water plumes or upwelling extents were deter

The extents of fresh water plumes or upwelling extents were determined by the positions of thermal fronts. These fronts were mapped by spatial domain filtration (3 × 3 window size) and calculating the gradient towards the local maximum of SST change, after previous median filtering to eliminate noise (Cayula and Cornillon, 1992 and Belkin and O’Reilly, 2009). The frontal zone was assumed to be an elongated, at least 5 km long, group of pixels with Veliparib molecular weight gradients over 0.2 °C km− 1. The project

commenced in 2008 and preliminary samples were collected from July to October. During this initial stage of the Ferry Box measurements a number of technical problems were encountered, one of the most annoying being severe fouling of the sensors by young forms of Mytilus trossulus and by Balanus spp.; malfunctioning of the WaterSam autosampler was also a common occurrence. The automatic measurements of temperature, salinity and chlorophyll

a showed a variability of environmental factors over the period from 11 July to 9 October 2008 ( Figure 2). Dissolved inorganic phosphate (DIP), oxygenated inorganic nitrogen (TO × N = NO3 + NO2), silicate, total phosphorus (TP) and total nitrogen (TN) were analysed in discrete seawater samples (TP and TN are not discussed here). Ammonia was not determined because the samples could not be treated with reagents NVP-BEZ235 immediately after sampling. Nutrient concentrations determined in discrete seawater samples depended on the station location and sampling date. Results from the off-shore station (GK4) are shown in Figure 3 to illustrate the observed fine changes in nutrient levels. From 7 July to 10 October 2008, chlorophyll a was measured in samples from discrete sampling stations on 11 occasions. The results showed considerable variability in chlorophyll

a concentrations, depending on the location of the sampling station and sampling date ( Figure 4). Phytoplankton species structure, abundance and biomass were determined in discrete samples on 3 occasions, between 7 and 28 July 2008 (Figure 5). The species structure showed considerable diversity (Figure 5). Flagellates were dominant in terms of both biomass and abundance, although there was also a marked presence of Dinophyceae in the biomass. The contributions of Cyanophyceae Nintedanib (BIBF 1120) and Bacillariophyceae to the biomass were considerable in the off-shore part of the ferry route. In fact, the biomass of the latter class consisted of a single diatom species – Actinocyclus octonarius. As for blue-green algae, the potentially toxic species Nodularia spumigena, accompanied by Aphanizomenon flosaquae, were dominant among the Cyanophyceae in general ( Figure 6), and Aphanothece paralleliformis was found to be dominant at a single station. The presence of nodularin was detected in discrete samples collected between 7 July and 13 August.

The soil column was placed in an 80 cm deep square

pit fi

The soil column was placed in an 80 cm deep square

pit filled with soil both inside and outside the column and made soil compact by watering. The distance between soil columns was 11 cm, that is, the row width was 65 cm, and surrounded by the board rows (Fig. 1). Two plants MS-275 in vivo were grown in each soil column. Plants in one column were planted under normal spacing (NS, 27 cm), and the other under narrow spacing (CS, 6 cm). The columns were treated at two nitrogen levels, N0 (no N) and N1 (7.5 g N plant− 1), and for the N1 treatment, nitrogen fertilizer was applied by 20%, 50% and 30% at the seedling, male-tetrad and flowering stages, respectively. The experimental design included four treatments MAPK inhibitor (N0 × NS, N0 × CS, N1 × NS and N1 × CS) and 30 separate soil columns were planted in each treatment. Samples of the soil columns (top 40 cm) were mixed and screened with 20 mesh sieving. Then they were mixed

with clean river sand in a ratio of 3:1 by volume of topsoil to sand. The mixed soil nutrient contents were as follows: organic matter 7.1 g kg− 1, total nitrogen (N) 0.62 g kg− 1, mean available mineral phosphorous (P) 46 mg kg− 1, and exchangeable potassium (K) 59 mg kg− 1. All treatments were fertilized with P and K according to nutrient demand, and each unit of experimental treatment was fertilized with 2.5 g of phosphate (P2O5) and 6.25 g of potash (K2O), with both applied at the seedling stage. Required irrigation was also applied from the outlet of a pump by using plastic pipes. At the onset of pollination, three replicates of each treatment were sampled on the same day fortnightly. The above-ground plant parts were divided into leaves, grains and stems (remaining parts except for leaves and grains). Roots were separated from various layers of the soil profile, viz. 0–20, 20–40, 40–70 and

> 70 cm, and washed to remove all soil residues. Root layers were mixed well after removing impurities, and fine roots were selected and temporarily stored at 0 °C. 2,3,5-triphenyl tetrazolium chloride (TTC) reduction was applied to determine root reductive activity [19]; fresh root samples (0.5 g) were exposed mafosfamide to 0.4% TTC and 0.2 mol L− 1 tricine-HCl buffer (pH 8.4), then placed in a darkroom at 37 °C for 6 h to induce reduction of TTC to triphenyl formazan (TTF), following the method described by Duncan and Widholm [19]. To quantify the amount of TTC reduced, we extracted the tissues with 95% ethanol at room temperature for 48 h, and then performed spectrophotometric analysis at 485 nm. The results were expressed as μg TTF g− 1 root mass h− 1. At maturity, the remaining aboveground parts were completely harvested to calculate average dry matter weight per plant and weight distribution. The remaining roots and above-ground samples were fixed at 105 °C for 30 min. Samples were subsequently baked at 75 °C until a constant weight was reached and recorded.