The second point will be discussed in the

following secti

The second point will be discussed in the

following section on management. Specific inputs are required for each type of development intervention (i.e., tourism, aquaculture, PES, etc.). A discussion of each livelihood is beyond the scope of the current paper; however, this review revealed a number of themes regarding the achievement of successful outcomes from various development interventions. First, the literature addresses how development needs to adopt participatory, adaptive, and equitable processes. Rarely are livelihoods initiatives imposed by organizations from the outside sustained over the long term. As an antidote to top-down development, participatory development processes may be more likely to lead to successful outcomes through facilitating co-learning and consensus-building, empowerment,

17-AAG cost and local mobilization [11], [76], [96], [104] and [127]. Simple processes, such as Participatory Rural Appraisal [165] or the Sustainable Livelihood Enhancement and Diversification (SLED) approach [159], can be used to facilitate participation Cisplatin mw in development. Development should also adopt an adaptive process of monitoring, feedback, and learning [35] and [111]. Adaptive learning also needs to be integrated into MPA-related conservation and development discourse and practice at a broader scale so that failed initiatives are not repeated and successes are recognized. Conservation and development programs should address the needs of potentially marginalized groups. Incorporating gender considerations, for example, into design of development programs and women׳s resource use patterns into MPA design can lead to greater benefits for households and the larger community [53], [78] and [93]. Participatory processes PDK4 can also lead to an improved understanding of the context from the perspective of local people which can be incorporated into the design of locally grounded and appropriate

solutions [104], [126] and [166]. Pre-assessments are important since assumptions about context can result in unsuccessful programs of action [167]. It is important to understand how micro to macro level contextual factors, such as access to markets, local capabilities, policy environments, levels of social cohesion, leadership capacity, and cultural norms, influence current marine uses and how these may facilitate or impede alternative livelihood development [35], [75], [161] and [168]. Third, authors suggest that development of alternative livelihoods often requires attention to building local capabilities through increasing financial and human capital, as well as physical assets (e.g., fishing gear, boats, basic and tourism infrastructure). Ongoing programs of education and capacity building are necessary for resource users to nurture occupational flexibility and acquire the skills necessary to engage in new livelihoods [17], [122], [160], [169] and [170].

Apoptosis was determined in cryosections obtained as described ab

Apoptosis was determined in cryosections obtained as described above from healthy vitellogenic and atretic follicles, using the ApopTag® Plus Apoptosis Detection Kit (Chemicon) PLX3397 cell line following manufacturer’s instructions, but extending the TdT incubation step to 16 h at 4 °C. Additional controls were also performed excluding the TdT enzyme from the labeling buffer and following the assay as above. Longitudinal sections were

revealed with DAB and photographed under light microscope. Yolk granule fractions from healthy vitellogenic and atretic follicles were obtained as described elsewhere (Ramos et al., 2007). The granules were incubated in the dark for 10 min in Ringer plus 10 mM EGTA containing 5 μg/ml acridine orange. After incubation the yolk granules were deposited on glass slides and observed in a Zeiss Axioplan epifluorescence microscope equipped with a fluorescein filter set and a TK-1270 JVC color video camera. Healthy vitellogenic and atretic follicles were dissected and homogenized on ice in phosphate buffer (0.1 M sodium phosphate, 0.2 M NaCl,

5 mM EDTA) pH 7.0 or acetate buffer (0.1 M sodium acetate, 0.2 M NaCl, 5 mM EDTA) pH 5.0. Ten follicles were used from each sample. Homogenates were submitted to three cycles of freeze and thaw and centrifuged at 20,000 × g see more for 30 min at 4 °C. Supernatants were collected and used as protease preparations. Protease assays were performed by incubating 0.1 follicle equivalents in 50 volumes of acetate buffer pH 4.0 plus 2.5 mM DTT and 10 μM Abz-AEALERMF-EDDnp (Aspartic), or acetate buffer pH 5.0 plus 2.5 mM DTT and 5 μM Z-Phe-Arg-NHMeC ID-8 (Serine and Cysteine). Substrate hydrolysis was monitored in an F-MAX 4500 fluorometer (Molecular Devices, Sunnyvale, CA, USA) at 320 nm excitation and 420 nm emission wavelengths for Abz-AEALERMF-EDDnp or 380 nm excitation and 440 nm emission wavelengths for Z-Phe-Arg-NHMeC.

Steady-state velocities were obtained by linear regression of the substrate hydrolysis curve ( Lima et al., 2001). Healthy vitellogenic and atretic follicles were centrifuged at 20,000 × g for 30 min at 4 °C. Supernatants were collected and used as samples for electrophoresis. Protein concentration was determined by the method of Lowry ( Lowry et al., 1951) using bovine serum albumin as standard. Polyacrylamide gels (10%) were run at 25 mA, applying 10 μg of protein per lane. Gels were silver stained using the protocol described by Dunn and Crisp (1994). Direct injection of conidia into the hemocoel of R. prolixus females at the onset of vitellogenesis did not affect host survival (Log-rank test, p = 0.5553, N = 31–33 subjects). Median survival was 23, 24.5 and 20 days for control (uninjected group); Grace’s injected group and fungal injected group, respectively, confirming the low pathogenicity of A. niger to these insects. Our previous results ( Medeiros et al., 2009) showed that R.

“Current Opinion in Behavioral

“Current Opinion in Behavioral find more Sciences 2015, 1:78–85 This review comes from a themed issue on Cognitive neuroscience Edited by Cindy Lustig and Howard Eichenbaum 2352-1546/© 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license ( It is an obvious, but sometimes overlooked, fact that it frequently takes many weeks to get an experimental animal to perform a task that could be explained to a human participant

in a matter of minutes. From one perspective, this neatly encapsulates how useful language is to communicate information. However, it also highlights just how important, and often difficult, it can be without such input to determine which specific elements of a complex environment should be used to guide and update behaviour. This is particularly evident in situations where stimuli and rewards are separated in space and time, can have different meanings depending on the external context or internal state, and can also provide several different types of information (for instance, a food or fluid reward might both satisfy an

internal need and provide information that the correct response has been made) [1]. One pressing question is Talazoparib therefore what neural structures help select relevant information and inhibit irrelevant information for the task in hand and how these relate to neural mechanisms implicated in value-guided decision making 2, 3, 4, 5, 6•• and 7]. A related issue concerns the mechanisms that allow us to determine, and potentially seek out, information relevant to satisfy

a current need, and also how these systems interrelate with circuits implicated in reward seeking [8]. While these are complex topics, in this brief review we will focus on converging evidence that the lateral parts of orbitofrontal cortex (OFC) and ventromedial prefrontal cortex (VMPFC) play key roles in these faculties. OFC and Methocarbamol VMPFC are large structures consisting of multiple distinct areas. Nonetheless, there are anatomical similarities between certain regions, which has allowed Price to define two distinct, though interconnected, networks in rodents, monkeys and humans [9]. First, an ‘orbital sensory’ network, including Walker’s areas 11, 12 and 13 and parts of anterior insula in primates, receives rich sensory information from all sensory modalities and also projects back to sensory structures. The equivalent network in the rat would include LO, VLO and AIv. By contrast, a ‘medial visceromotor’ network, including medial OFC area 14 as well as areas 25 and 32 and medial area 10, is characterised by strong connections with the medial temporal lobe as well as projections to limbic regions such as ventral striatum and lateral hypothalamus. In the rat, this network is likely made up of MO (medial orbital), prelimbic and infralimbic cortex.

The overall effect on spinal neuronal activity is dependent on th

The overall effect on spinal neuronal activity is dependent on the interplay between excitatory and inhibitory mechanisms; thus, our data suggest that the overriding effects of ketanserin and ritanserin were

likely to be mediated through antagonism of the actions of 5-HT acting at 5-HT2A receptors leading to the reduction in neuronal responses observed in this study. The consequence of 5-HT2C receptor blockade, at these doses, on the evoked spinal neuronal responses is minimal by comparison, if 5-HT2C receptors do indeed have an antinociceptive role. Similarly, activation of 5-HT2A/2C receptors with DOI increased neuronal responses, click here an effect reversed by ketanserin, thus implicating a predominant 5-HT2A action. An alternative possibility, however, is that 5-HT2C receptors could also have pronociceptive effect on spinal nociceptive transmission. The primary source LDE225 research buy of descending serotonergic modulation of ascending nociceptive transmission from the spinal cord arises from the RVM (Millan, 2002). These serotonergic neurones can exert facilitatory or inhibitory influences onto dorsal horn neurones depending on the spinal 5-HT receptor subtype activated and the neuronal cell type within the RVM (Millan, 2002). Neurones within the RVM are classified into three types based upon their firing patterns in response to noxious thermal stimuli. ON-cells increase their firing immediately before a nocifensive

response and facilitate

nociception, while OFF-cells, considered to mediate inhibition, pause in their firing just prior to a nociceptive withdrawal reflex. Neutral cells do not appear to play a role in physiological pain (Heinricher et al., 2009). Descending facilitation requires the activation of pronociceptive ON cells (Porreca et al., 2001); however, the pharmacology of descending facilitatory pathways remains unclear, as recordings from RVM neurones suggests that 5-HT containing neurones Montelukast Sodium are neither ON nor OFF cells (Gao and Mason, 2000). However, converging evidence from recent immunohistochemical, behavioural and electrophyisological data suggests that a proportion of RVM cells activated by noxious stimuli are serotonergic. Furthermore, a facilitatory effect mediated by 5-HT, acting at spinal 5-HT3 receptors, was demonstrated in models of acute and chronic pain (Oatway et al., 2004, Rahman et al., 2004, Rahman et al., 2006, Suzuki et al., 2002, Suzuki et al., 2005 and Svensson et al., 2006). These studies focused on the pronociceptive 5-HT3 receptor, the only ligand gated cation channel of the 5-HT receptor family, and its role in mediating descending facilitation. The electrophysiological consequences of selectively blocking spinal 5-HT2A receptors on dorsal horn neuronal activity are similar to the effects we have previously seen with the selective 5-HT3R antagonist ondansetron (Suzuki et al., 2002).

We obtained many aerobic cellulolytic microorganisms which were d

We obtained many aerobic cellulolytic microorganisms which were distinguished based on their colony morphology. Among them, a bacterial isolate JS-C42 exhibited highest lignocellulolytic effect. In this study,

we are presenting a detailed report of a yellow actinomycete Dabrafenib order isolate JS-C42. Plating of the cultures of JS-C42 on cellulose agar during subsequent sub culturing also depicted the extensive clearing zones. The clearing zone shown depicted the cellulose solubilization by extracellular enzymes produced by JS-C42 isolate and this result was in accordance with the cellulolytic studies as reported by Sizova et al. [24]. Cellulolytic strain JS-C42 has a smooth surface, pale yellow, circular, opaque colonies and approximately 1.0 mm in diameter after 36 h growth at 28 °C on cellulose supplemented medium. It grew well at pH 7.5–9.0, 28–37 °C and up to 10% NaCl concentration. The cells were Gram-positive, non-motile cocci-shaped, have primary mycelium with no spore and exhibited aerobic growth. The bacterial isolate JS-C42 utilized the starch, casein, urea and lipid molecule such as tributyrin.

The utilization of starch, casein, mannitol salt agar, tributyrin, and urea showed that the isolate produced the extra cellular enzymes amylase, protease, lipase and urease to metabolize the polymeric components of the nutrient mixture to monomeric form for the growth. For predicting the Belnacasan phylogenetic position of the isolate JS-C42, the phylogenetic tree (Fig. 1) with its closely related type and non-type strains were analyzed using Ribosomal Database Project. The nucleotide sequence of the 16S rRNA gene of JS-C42 displayed 98.9% sequence identity to the available 16S rRNA gene sequence of the type strain Isoptericola halotolerans YIM 70177 Chloroambucil and 99.3% sequence similarity to the non type strain Isoptericola sp. DSX2. The closely related type strain Isoptericola halotolerans YIM 70177 was negative for milk peptonization and starch hydrolysis and its colonies are pale-yellow in color [25]. When compared to the type strain Isoptericola halotolerans YIM 70177

and in spite of the 16S rRNA gene sequence identity of 98.9%, the cellulolytic bacterial isolate JS-C42 showed phenotypic differences in cell morphology like intense yellow with distinct mycelium and distinct biochemical properties like positive reaction for milk peptonization and starch hydrolysis. Overall the phylogenetic analysis of cellulolytic bacterial isolate JS-C42 revealed its belongings to the phylum Actinomyces and denoted as Isoptericola sp. JS-C42. The cellulose hydrolysis is observed after the 48 h incubation with a zone of the hydrolyzed region of the cellulosic agar medium flooded with Gram’s iodine, which produces a bluish-black complex with cellulose but not with hydrolyzed zone containing simple sugars [10].

3) Dynorphin1-13 (dyn A, YGGFLRRIRPKLK) was also hydrolyzed

3). Dynorphin1-13 (dyn A, YGGFLRRIRPKLK) was also hydrolyzed Panobinostat solubility dmso by the crude venom of B. jararaca, showing at least two cleavage points (YGGFLR-RIRPK-LK), since the fragment RIRPK was detected by mass spectrometry analyses. Unlike angiotensin I, dyn A is hydrolyzed by both classes of proteases, metallo- and serine peptidases, so this activity was partially blocked by the commercial antibothropic serum. The pathophysiological mean of dyn A hydrolyzes is possibly correlated with pain sensation and inflammation ( Parikh et al., 2010 and Luo et al., 2008). Many factors,

including phylogeny, sex, geographic origin, season, age and prey preference, Cytoskeletal Signaling inhibitor may influence composition of the venoms (Chippaux et al., 1991, Mackessy et al., 2003 and Furtado et al., 2006). In addition to these considerations, the genus Bothrops shows the greatest diversity when it comes to number of species, morphology and natural history characteristics ( Campbell and Lamar, 2004). Given these characteristics, the development of a polyvalent antivenom against accidents involving this genus is an even greater challenge. Thus, the production

of better antivenoms should take into consideration the quality of poisons, and what poisons should be used to compose the pool of immunization. Finally, the preclinical efficacy of the antivenom must be carefully evaluated. The inter specimen venom composition may be evidenced by the different levels of chymotrypsin-like activity and by the different potential GPX6 blockers obtained with the antibothropic serum and the five different Bothrops venoms studied in this paper. These venom composition variations may be an important factor to explain the failure of the antibothropic serum and, additionally, three other factors also may be responsible for the overall presented result. The first factor suggests a lack of immunoglobulins acting against serine peptidases present in some venoms and

the second factor may be related to the failure of blocking by the antibodies, although they may be present. The third and important factor may be related to degradation of the serine peptidases by the metallo peptidases before the inoculation of horses with the pool of venoms used for the production of antivenom, and this degradation could destroy the epitopes responsible for the production of immunoglobulins. These hypotheses are under investigation in our laboratories through new experiments, with the objective of developing strategies to obtain a more effective antibothropic serum. The antibothropic serum produced by the Butantan Institute is one of the best in Latin America to reduce mortality by snake poisoning from this genus.

Identification of key events at the transcriptional level can fac

Identification of key events at the transcriptional level can facilitate the identification of processes that are critical for disease initiation and progression, thus allowing information from animal experiments to be queried and used for extrapolation to human scenarios (Edwards and Preston, 2008). Comparison of our data with specific models of lung disease, including bacterial infection, airway hypersensitivity and lung injury revealed that CBNPs induced Ibrutinib price responses that were more closely related to lung injury and fibrosis than to other models. This finding was further supported

by comparison of the expression profiles of CBNP exposed mice to those of curated studies of animals and humans exhibiting a myriad of pulmonary disease phenotypes. This analysis demonstrates that CBNP exposure perturbs genes that are Dasatinib supplier known to be involved in tissue injury and fibrosis in mice. Although it is unclear if CBNP exposure would result in the same gene expression profile

in humans, similar pathways including many involved in fibrotic responses were found in both mice and humans (52% of the top 50 pathways found were common between mouse and human). Despite concordance of pathways, the top ranked genes differed considerably between both species. However, many of the genes found in mice and humans had similar functions, including inflammatory and acute phase responses (e.g., Saa3, Socs3 and Mt2 in mice and CP, VNN2 and CXCL10 in humans), cell cycle progression (Cdkn1a in mice and KLF4 in humans) and bone and tissue modelling

(Mmp14, Timp1, Eln and Ogn in mice and SPP1 in humans). Thus, despite discordance in the gene expression profiles between species, the similar functions Oxymatrine of top ranked genes and concordance between pathways supports the likelihood of similar responses in the event of CBNP exposure in humans. In addition, fibrosis has been identified as an outcome of exposure to various particles and NPs in animals ( Bermudez et al., 2004 and Shvedova et al., 2008), including Printex 90 (e.g., 28-day nose only inhalation in Wistar WU rats) ( Bellmann et al., 2009), as well as in humans ( Lkhasuren et al., 2007 and Wang and Christiani, 2003). The process of pulmonary fibrosis is closely related to progression of carcinogenic outcome ( Hubbard et al., 2000). These data demonstrating very similar fibrotic pathways in mice and humans and a significant overlap with CBNP-induced gene expression changes thus support the use of pathway-based approaches in identifying molecular mechanisms of disease onset and progression, and using gene expression profiles to support HHRA. This study confirms several key elements that are necessary for the application of gene expression profiling for HHRA of toxicant exposures in general. First, transcriptional profiles can effectively predict the biological effects of chemical exposures.

The resonant frequency of water depends on the

The resonant frequency of water depends on the Selleck EPZ015666 temperature. The temperature dependence of the resonant frequency of 1H in water is about 0.01 ppm/°C [18]. The temperature of MEA may rise due to heat generation in the PEFC and, as a result, the resonance frequency of 1H of water in MEA may change. When this change due to temperature rise is large, the assumption that resonance frequency changes only due to magnetic fields induced by electric current within the PEFC is not valid. When the PEFC employed generates a current of 5 A, the heat generation of the PEFC is estimated to be about 2 W. The temperature rise of the MEA due to a heat generation of 2 W is further estimated to be about 1 °C at the

most from a heat transfer analysis. When the temperature of the MEA rises by 1 °C, the change of the resonance frequency of 1H is about 0.01 ppm. On the other hand, when the PEFC used here generates an electric current of 5 A, the fluctuation of the frequency check details shift obtained from NMR signal mixed with noise is about 7–10% of the frequency shift. The corresponding variation of the measured frequency shift is from 0.7 to 5.5 ppm. Therefore, we think that the change of the resonant frequency of 1H (water) due to temperature rise of MEA hardly affects the calculation

of electric current generated in the PEFC. We can understand the electrical generation and the time dependent change of the water which has formed inside the PEFC by simultaneously measuring the spatial distributions of the water content in the PEM and the local current density within the PEFC. We expect that the system developed here will prove useful in the research into suitable control procedures and appropriate PEFC structures to allow the stable generation of electrical power in PEFCs. In order to measure the time-dependent change of the spatial distributions of current density and water content in a PEM, we have developed an eight-channel NMR system. Eight RF detection coils of 0.6 mm inside diameter were inserted in the PEFC at different positions. The Carbohydrate NMR signals from water in the PEM at these eight positions were then acquired simultaneously.

The spatial distribution of current density generated in the PEFC and the water content in the PEM could be calculated from the frequency shift and the amplitude of the obtained NMR signal. The NMR system was developed by MRTechnology, Inc., NEOMAX Engineering, Ltd. and Digital Signal Technology, Inc. The software for the NMR measurements was programmed by Mr. Seitaro Hashimoto of EXA CORPORATION. The MEA was built by Dr. Sangkun Lee and Mr. Masaaki Hirano of the Hydrogen Utilization Engineering Kyusyu University. Some parts of the fuel cells were made by FC composite Inc. and Yamato Inc. The authors wish to thank all of those mentioned above for their contributions to this study. “
“In Fig. 4 the denomination of the regions for the substances 1, 2, 3 and 5 has to be changed to that shown in the corrected figure.

It is not clear if the model described by Ma et al (2013) overes

It is not clear if the model described by Ma et al. (2013) overestimates wave height or Fluidity underestimates. It should be noted that previous comparisons of Fluidity to both numerical models and observational data, Haugen et al. (2005) and Oishi et al. (2013), show excellent

agreement to both amplitude and phase of wave patterns resulting from both slides and earthquakes in two- and three-dimensions at ocean scales. Having benchmarked the implementation of the prescribed slide boundary conditions against independent models, we now show how Fluidity is capable of simulating real-world scale slide-generated tsunamis with high resolution in areas of interest by recreating the Storegga slide. The same domain is used

for all simulations described here. The domain stretches from 43° west to 24° east and 47° north to selleck chemical 80° north. GSHHS data (Wessel and Smith, 1996) was used to generate coastlines for all modern simulations, which has resolutions of 200 m (full) to 25 km (coarse). For the simulation involving palaeobathymetry the coastline was derived from the 0 m contour. Bathymetric data was derived from GEBCO (IOC, 2008) which has resolution of 1 arcminute (approximately 2 km in this region). For each domain QGIS (QGIS Development Team, 2009) was used with bespoke software to generate coastline input for GMSH (Geuzaine and Remacle, 2009) which created the horizontal computational mesh. The mesh is on a Cartesian sphere of radius 6371.01 km. Coastlines were constructed using a B-spline

curve through the points given by the GSHHS data. Bathymetry is incorporated by extruding the generated surface mesh radially downward to the depth given by the bathymetric data, which is carried out at run-time. Each simulation uses a one-element deep solution, Resminostat effectively a depth-averaged velocity as used in (Mitchell et al., 2010 and Wells et al., 2010). A consequence of this approximation is that a minimum water depth has to be specified for the mesh as inundation (wetting and drying) was not utilised in this study. Here, a minimum depth of 10 m was used. We generate the slide using the single rigid block slide, described in Eqs. (4), (5), (6), (7), (8), (9), (10) and (11), following the work in Harbitz (1992), using the parameters in Table 2. Note that we do not include the effects of retrogressive slide evolution. This style of multi-block slide motion was investigated in Løvholt et al. (2005) and Bondevik et al. (2005), who concluded that the time interval between block initiation would need to be very small in order to produce large wave heights consistent with observation and such scenarios are qualitatively similar to the motion of a single continuous body. For initial runs, to explore the sensitivity of model results to spatial resolution, the simulation was run for five hours model time, which was sufficient to allow comparison with previous studies.

To generate

a BSMV:TaWAK5 construct, a 298 bp sequence of

To generate

a BSMV:TaWAK5 construct, a 298 bp sequence of TaWAK5 (from nucleotide position 1913 to 2211 in the TaWAK5 cDNA sequence) was amplified from the cDNA sequence for TaWAK5 buy Ion Channel Ligand Library from the genotype CI12633 with the primers TaWAK5-VIGS-F/TaWAK5-VIGS-R. PCR-amplified cDNA fragments were digested with Pac I and Not I, then ligated into the BSMV:RNAγ vector digested with Pac I-Not I, resulting in the recombinant construct RNAγ:TaWAK5-as. Following a previously described protocol [32], the tripartite cDNA chains of BMSV:TaWAK5, or the control virus BMSV:GFP genome, were separately transcribed into the RNAs, then mixed and used to infect CI12633 plants at the 2-leaf stage. At the same time, CI12633 plants were inoculated with only the buffer without virus. Hereafter, these plants treated only with buffer are referred to as mock treatments. The 4th leaves of the inoculated seedlings were collected and analyzed for the virus infection based on the RNA transcript presence of the BSMV coat protein gene using Ku-0059436 cost primers BSMV-CP-F/BSMV-CP-R. These tissues were also evaluated for changes in TaWAK5 expression with primers TaWAK5-Q-F/TaWAK5-Q-R

at 10 days after BSMV infection. For R. cerealis inoculation, the fungus was cultured on potato dextrose agar at 25 °C for 10 days, then 1 cm2 plugs from the edge of R. cerealis colonies were placed into liquid PDA medium and cultured at 25 °C for 2 weeks, to develop the mycelia. The 4th base sheath of wheat plants was inoculated with 15 μL of the R. cerealis liquid culture at 20 days after BSMV virus inoculation. Inoculated plants were grown at 90% relative humidity for 4 days. Sharp eyespot symptoms were observed respectively at 14 days and 40 days after fungal inoculation. These are the times when sharp eyespot symptoms are normally present at the infected sheaths and stems, respectively, of the susceptible cultivar Wenmai 6. RT-PCR was performed with 20 μL reaction volumes from the TaKaRa Inc. kit containing 1 × PCR buffer, 2.0 μL 10 × first strand cDNA,

150 μmol L− 1 of each dNTP, and 1 U Taq polymerase, plus 0.25 μmol L− 1 of each primer. The program used was as follows: initial denaturation at 94 °C for 5 min; followed by 30 cycles Astemizole of 30 s at 94 °C, 30 s at 60 °C, and 30 s at 72 °C; and final extension at 72 °C for 5 min. The PCR products were detected on 2% agarose gels. In all the semi-quantitative RT-PCR experiments, wheat elongation factor 1 alpha-subunit (TaEF-1a) was used to normalize the cDNA contents among various samples. qRT-PCR was performed using SYBR Green I Master Mix from TaKaRa Inc. in a volume of 25 μL on an ABI 7300 RT-PCR system (Applied Biosystems Corp.). Reactions were set up with the following thermocycling profile: 95 °C for 5 min, followed by 41 cycles of 95 °C for 15 s and 60 °C for 31 s. The products were continuously examined with a melting curve analysis program.