Biostimulants induce positive changes in the radish morpho-physiology and yield.

An ever-increasing population has issued an open challenge to the agricultural sector to provide enough food in a sustainable manner. The upsurge in chemical fertilizers to enhance food production had resulted in environmental problems. The objective of the current study is to assess the utilization of biostimulants for sustainable agricultural production as an alternative to chemical fertilization. For this purpose, two pot experiments were conducted to examine the response of radish against individual and combined applications of biostimulants. In the first experiment, the effects of chemical fertilizer (CK), glycine (G), lysine (L), aspartic acid (A), and vitamin B complex (V) were studied. The results demonstrated that V significantly improved the transpiration rate (81.79%), stomatal conductance (179.17%), fresh weight (478.31%), and moisture content (2.50%). In the second experiment, tested treatments included chemical fertilizer (CK), Isabion® (I), glycine + lysine + aspartic acid (GLA), moringa leaf extract + GLA (M1), 25% NPK + M1 (M2). The doses of biostimulants were 5g L-1 glycine, 1g L-1 lysine, 2g L-1 aspartic acid, and 10 ml L-1 moringa leaf extract. The photosynthetic rate improved significantly with GLA (327.01%), M1 (219.60%), and M2 (22.16%), while the transpiration rate was enhanced with GLA (53.14%) and M2 (17.86%) compared to the Ck. In addition, M1 increased the stomatal conductance (54.84%), internal CO2 concentration (0.83%), plant fresh weight (201.81%), and dry weight (101.46%) as compared to CK. This study concludes that biostimulants can effectively contribute to the sustainable cultivation of radish with better growth and yield.

Application of K and Zn Influences the Mineral Accumulation More in Hybrid Than Inbred Maize Cultivars.

Maize (Zea mays L.) is an important crop used for feeding humans and cattle globally. Deficiency of potassium (K) and zinc (Zn) adversely impacts the maize crop productivity and quality. However, the application of these nutrients shows variant responses in different maize cultivars. To understand this perspective, the current study aimed at investigating K and Zn's optimal concentration in different hybrid and inbred maize cultivars. The treatments were based on three zinc levels (0, 6, and 12 mg Zn kg-1) and K levels (0, 30, and 60 mg kg-1), and their respective combinations. The experiment results showed that combined fertilization approaches of Zn and K (Zn12K60) improved the plant biometric, and physiological attributes of maize crop. The results revealed a significant increase in plant height (45%), fresh weight (70%), and dry weight (45%). Similarly, physiological attributes significantly improved the relative water content (76.4%), membrane stability index (77.9%), chlorophyll contents (170%), and photosynthetic rate (130%) in both inbred and hybrid genotypes. Furthermore, Zn and K (Zn12K60) increased transpiration rate (E), stomatal conductance (Ci), and internal CO2. In conclusion, maize hybrids (Neelam and DK-6142) were observed best compared with inbred (Afghoi and P-1543) cultivars with the combined application of Zn and K (Zn12K60). Thus, these inbred varieties should be preferred for fodder requirement with optimum fertilizer (Zn12K60) application in Zn deficient soils.

Synthesis and Identification of a Novel Peptide, Ac-SDK (Biotin) Proline, That Can Elicit Anti-Fibrosis Effects in Rats Suffering from Silicosis.

BACKGROUND: N-Acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) is a short peptide with an anti-silicosis effect. However, the short biological half-life and low plasma concentration of Ac-SDKP hamper discovery of specific targets in organisms and reduce the anti-silicosis effect. A novel peptide, Ac-SDK (biotin) proline, termed "Ac-B", with anti-fibrotic properties was synthesized. METHODS: Ac-B was detected quantitatively by high-performance liquid chromatography. Phagocytosis of Ac-B by the alveolar epithelial cell line A549 was investigated by confocal laser scanning microscopy and flow cytometry. To further elucidate the cellular-uptake mechanism of Ac-B, chemical inhibitors of specific uptake pathways were used. After stimulation with transforming growth factor-ß1, the effects of Ac-B on expression of the myofibroblast marker vimentin and accumulation of collagen type I in A549 cells were analyzed by Western blotting. Sirius Red staining and immunohistochemical analyses of the effect of Ac-B on expression of α-smooth muscle actin (SMA) in a rat model of silicosis were undertaken. RESULTS: Ac-B had good traceability during the uptake, entry, and distribution in cells. Ac-B treatment prevented an increase in α-SMA expression in vivo and in vitro and was superior to that of Ac-SDKP. Caveolae-mediated uptake of Ac-B by A549 cells led to achieving anti-epithelial-mesenchymal transformation (EMT) effects. CONCLUSION: Ac-B had an anti-fibrotic effect and could be a promising agent for the fibrosis observed in silicosis in the future.

N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) attenuates silicotic fibrosis by suppressing apoptosis of alveolar type II epithelial cells via mediation of endoplasmic reticulum stress.

Damage to alveolar epithelial cells (AECs) caused by long-term inhalation of large amounts of silica dust plays a significant role in the pathology of silicosis. The present study was undertaken to investigate the regulatory mechanism(s) involved in type II AEC damage from silicon dioxide (SiO2) as well as the mechanism(s) related to the prevention of silicosis by the antifibrotic tetra peptide, N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP). The 2-DE results showed that SiO2 induced endoplasmic reticulum (ER) stress in A549 cells. In addition, typical apoptotic characteristics were observed using a transmission electron microscope (TEM) in A549 cells stimulated by SiO2 and in type II AECs from silicotic rats. Mechanistic study showed that both Ac-SDKP and 4-phenylbutyrate (4-PBA), an inhibiter of ER stress, attenuated GRP78, phosphor-PERK, phosphor-eIF2α, CHOP and Caspase-12 protein expression in A549 cells stimulated by SiO2 and in type II AECs from silicotic rats. Treatment with Ac-SDKP and 4-PBA in vivo effectively inhibited collagen deposition in the lungs of silicotic rats. In summary, ER stress is involved in the apoptosis of type II AECs both in vitro and in vivo. Ac-SDKP effectively suppresses SiO2-induced apoptosis in type II AECs by attenuating the Caspase-12 and PERK/eIF2α/CHOP pathway activation caused by ER stress, thus preventing silicotic fibrosis.

Influence of the interaction between Ac­SDKP and Ang II on the pathogenesis and development of silicotic fibrosis.

N­acetyl­seryl­aspartyl­lysyl­proline (Ac­SDKP) is a natural tetrapeptide that is released from thymosin ß4 by prolyl oligopeptides. It is hydrolyzed by the key enzyme of the renin­angiotensin system, angiotensin­converting enzyme (ACE). The aim of the present study was to investigate the alterations in Ac­SDKP and the ACE/angiotensin II (Ang II)/angiotensin II type 1 (AT1) receptor axis and its impact on the pathogenesis and development of silicotic fibrosis. For in vivo studies, a HOPE MED 8050 exposure control apparatus was used to establish different stages of silicosis in a rat model treated with Ac­SDKP. For in vitro studies, cultured primary lung fibroblasts were induced to differentiate into myofibroblasts by Ang II, and were pretreated with Ac­SDKP and valsartan. The results of the present study revealed that, during silicosis development, ACE/Ang II/AT1 expression in local lung tissues increased, whereas that of Ac­SDKP decreased. Ac­SDKP and the ACE/AT1/Ang II axis were inversely altered in the development of silicotic fibrosis. Ac­SDKP treatment had an anti­fibrotic effect in vivo. Compared with the silicosis group, the expression of α­smooth muscle actin (α­SMA), Collagen (Col) I, Fibronectin (Fn) and AT1 were significantly downregulated, whereas matrix metalloproteinase­1 (MMP­1) expression and the MMP­1/tissue inhibitor of metalloproteinases­1 (TIMP­1) ratio was increased in the Ac­SDKP treatment group. In vitro, pre­treatment with Ac­SDKP or valsartan attenuated the expression of α­SMA, Col I, Fn and AT1 in Ang II­induced fibroblasts. In addition, MMP­1 expression and the MMP­1/TIMP­1 ratio were significantly higher in Ac­SDKP and valsartan pre­treatment groups compared with the Ang II group. In conclusion, the results of the present study suggest that an imbalance between Ac­SDKP and ACE/Ang II/AT1 molecules promotes the development of silicosis and that Ac­SDKP protects against silicotic fibrosis by inhibiting Ang II­induced myofibroblast differentiation and extracellular matrix production.

Long-term manure application increased greenhouse gas emissions but had no effect on ammonia volatilization in a Northern China upland field.

The impacts of manure application on soil ammonia (NH3) volatilization and greenhouse gas (GHG) emissions are of interest for both agronomic and environmental reasons. However, how the swine manure addition affects greenhouse gas and N emissions in North China Plain wheat fields is still unknown. A long-term fertilization experiment was carried out on a maize-wheat rotation system in Northern China (Zea mays L-Triticum aestivum L.) from 1990 to 2017. The experiment included four treatments: (1) No fertilizer (CK), (2) single application of chemical fertilizers (NPK), (3) NPK plus 22.5t/ha swine manure (NPKM), (4) NPK plus 33.7t/ha swine manure (NPKM+). A short-term fertilization experiment was conducted from 2016 to 2017 using the same treatments in a field that had been abandoned for decades. The emissions of NH3 and GHGs were measured during the wheat season from 2016 to 2017. Results showed that after long-term fertilization the wheat yields for NPKM treatment were 7105kg/ha, which were higher than NPK (3880kg/ha) and NPKM+ treatments (5518kg/ha). The wheat yields were similar after short-term fertilization (6098-6887kg/ha). The NH3-N emission factors (EFamm) for NPKM and NPKM+ treatments (1.1 and 1.1-1.4%, respectively) were lower than NPK treatment (2.2%) in both the long and short-term fertilization treatments. In the long- and short-term experiments the nitrous oxide (N2O) emission factors (EFnit) for NPKM+ treatment were 4.2% and 3.7%, respectively, which were higher than for the NPK treatment (3.5% and 2.5%, respectively) and the NPKM treatment (3.6% and 2.2%, respectively). In addition, under long and short-term fertilization, the greenhouse gas intensities for the NPKM+ treatment were 33.7 and 27.0kg CO2-eq/kg yield, respectively, which were higher than for the NPKM treatment (22.8 and 21.1kg CO2-eq/kg yield, respectively). These results imply that excessive swine manure application does not increase yield but increases GHG emissions.

Ammonia emissions from paddy fields are underestimated in China.

Excessive nitrogen (N) fertilizers are often used in China, and a large proportion of the N can be lost as ammonia (NH3). However, quantifying the NH3 emission from paddy fields is always affected by large uncertainties due to different measuring methods and other factors such as climate. In this study, using a standardized method, we measured the NH3 emissions in three typical annual rice cropping systems: single rice, double rice and rotation with other crops. The measurements were conducted for 2 years with a total of 3131 observations across China. Results showed that NH3 emissions accounted for 17.7% (14.4-21.0%) of the N applied under current farm practice, which was 33.1% (10.6-52.6%) higher than previous estimates. Nitrogen application rate was the dominant factor influencing NH3 emission rate, which exponentially increased with the N fertilizer rate (p < .001). Total NH3 emissions from paddy fields were estimated at 1.7 Tg N yr-1 in 2013 in China, several times the amount of N lost through leaching or runoff. This suggests that mitigation measures for non-point source pollution from cropland should take into account not only the N lost to water, but also to air, thereby improving air quality.

Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis.

BACKGROUND: Myofibroblasts play a major role in the synthesis of extracellular matrix (ECM) and the stimulation of these cells is thought to play an important role in the development of silicosis. The present study was undertaken to investigate the anti-fibrotic effects of dibutyryl-cAMP (db-cAMP) on rats induced by silica. METHODS: A HOPE MED 8050 exposure control apparatus was used to create the silicosis model. Rats were randomly divided into 4 groups: 1)controls for 16 w; 2)silicosis for 16 w; 3)db-cAMP pre-treatment; 4) db-cAMP post-treatment. Rat pulmonary fibroblasts were cultured in vitro and divided into 4 groups as follows: 1) controls; 2) 10-7mol/L angiotensin II (Ang II); 3) Ang II +10-4 mol/L db-cAMP; and 4) Ang II + db-cAMP+ 10-6 mol/L H89. Hematoxylin-eosin (HE), Van Gieson staining and immunohistochemistry (IHC) were performed to observe the histomorphology of lung tissue. The levels of cAMP were detected by enzyme immunoassay. Double-labeling for α-SMA with Gαi3, protein kinase A (PKA), phosphorylated cAMP-response element-binding protein (p-CREB), and p-Smad2/3 was identified by immunofluorescence staining. Protein levels were detected by Western blot analysis. The interaction between CREB-binding protein (CBP) and Smad2/3 and p-CREB were measured by co-immunoprecipitation (Co-IP). RESULTS: Db-cAMP treatment reduced the number and size of silicosis nodules, inhibited myofibroblast differentiation, and extracellular matrix deposition in vitro and in vivo. In addition, db-cAMP regulated Gαs protein and inhibited expression of Gαi protein, which increased endogenous cAMP. Db-cAMP increased phosphorylated cAMP-response element-binding protein (p-CREB) via protein kinase A (PKA) signaling, and decreased nuclear p-Smad2/3 binding with CREB binding protein (CBP), which reduced activation of p-Smads in fibroblasts induced by Ang II. CONCLUSIONS: This study showed an anti-silicotic effect of db-cAMP that was mediated via PKA/p-CREB/CBP signaling. Furthermore, the findings offer novel insight into the potential use of cAMP signaling for therapeutic strategies to treat silicosis.

Isopropanol biodegradation by immobilized Paracoccus denitrificans in a three-phase fluidized bed reactor.

A three-phase bed bioreactor including a mix of immobilized microbes was used to degrade isopropanol (IPA). The immobilization method was studied and cells immobilized with calcium alginate, polyvinyl alcohol, activated carbon, and SiO2 were demonstrated to be the best immobilization method for the degradation of 90% of 2 g/L IPA in just 4 days, 1 day earlier than with free cells. Acetone was monitored as an indicator of microbial IPA utilization as the major intermediate of aerobic IPA biodegradation. The bioreactor was operated at hydraulic retention time (HRT) values of 32, 24, 16, 12, and 10 hr, which correspond to membrane fluxes of 0.03, 0.04, 0.06, 0.08, and 0.10 L/m2/hr, respectively. The chemical oxygen demand (COD) removal efficiencies were maintained at 98.0, 97.8, 89.1, 80.6, and 71.1% at a HRT of 32, 24, 16, 12, and 10 hr, respectively, while the IPA degradations were 98.6, 98.3, 90.3, 81.6, and 73.3%, respectively. With a comprehensive consideration of COD removal and economy, the optimal HRT was 24 hr. The results demonstrate the potential of immobilized mixed bacterial consortium in a three-phase fluidized bed reactor system for the aerobic treatment of wastewater containing IPA.

Biodegradation of isopropanol by a solvent-tolerant Paracoccus denitrificans strain.

The biodegradation of high concentration isopropanol (2-propanol, IPA) at 16 g/L was investigated by a solvent-tolerant strain of bacteria identified as Paracoccus denitrificans for the first time by 16S rDNA gene sequencing. The strain P. denitrificans GH3 was able to utilize the high concentration of IPA as the sole carbon source within a minimal salts medium with a cell density of 1.5×10(8) cells/mL. The optimal conditions were found as follows: initial pH 7.0, incubation temperature 30°C, with IPA concentration 8 g/L. Under the optimal conditions, strain GH3 utilized 90.3% of IPA in 7 days. Acetone, the major intermediate of aerobic IPA biodegradation, was also monitored as an indicator of microbial IPA utilization. Both IPA and acetone were completely removed from the medium following 216 hr and 240 hr, respectively. The growth of strain GH3 on IPA as a sole carbon and energy source was well described by the Andrews model with a maximum growth rate (µmax)=0.0277/hr, a saturation constant (KS)=0.7333 g/L, and an inhibition concentration (Ki)=8.9887 g/L. Paracoccus denitrificans GH3 is considered to be well used in degrading IPA in wastewater.

Biodegradation of cypermethrin by a novel Catellibacterium sp. strain CC-5 isolated from contaminated soil.

The bacterial strain CC-5, isolated from contaminated soil and identified as Catellibacterium sp. based on morphology and partial 16S rDNA gene sequence analysis, utilized cypermethrin as its sole carbon source and degraded 97% of 100 mg·L(-1) cypermethrin within 7 days. The optimal degradation conditions were determined to be 30 °C and pH 7.0. Degradation was found to follow a first-order model at initial cypermethrin concentrations below 400 mg·L(-1). Strain CC-5 suffered substrate inhibition at high cypermethrin concentrations, and the biodegradation kinetics were successfully described by the Haldane model, with a maximal specific degradation rate of 1.36 day(-1), an inhibition constant of 164.61 mg·L(-1), and a half-saturation constant of 101.12 mg·L(-1). Inoculating cypermethrin-treated soil samples with strain CC-5 resulted in a higher rate of cypermethrin removal than that in noninoculated soil, regardless of whether the soil had previously been sterilized. These results reveal that the bacterial strain may possess potential to be used in bioremediation of pyrethroid-contaminated environment.

Isolation, identification and characterization of a glyphosate-degrading bacterium, Bacillus cereus CB4, from soil.

A bacterial strain named CB4, with highly effective glyphosate degradation capability, was isolated from soil after enrichment. On the basis of the Biolog omniLog identification system (Biolog) and 16S ribosomal RNA (rRNA) gene sequencing methods, strain CB4 was identified as Bacillus cereus. Further experiments were carried out to optimize the growth of strain CB4 and the glyphosate degradation activity by high performance liquid chromatography (HPLC). The optimal conditions were found as follows: initial pH 6.0, incubation temperature 35°C, glyphosate concentration 6 g L(-1), inoculation amount 5% and incubation time 5 days. Under the optimal conditions, stain CB4 utilized 94.47% of glyphosate. This is the first report on B. cereus with a capacity to utilize herbicide glyphosate, and it can degrade glyphosate concentrations up to 12 g L(-1). Metabolization of glyphosate by strain B. cereus CB4 was studied. Results indicated that two concurrent pathways were capable of degrading glyphosate to AMPA, glyoxylate, sarcosine, glycine and formaldehyde as products. Glyphosate breakdown in B. cereus CB4 was achieved by the C-P lyase activity and the glyphosate oxidoreductase activity.