Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

Thermal,swelling and stability kinetics of chitosan based semi-interpenetrating network hydrogels

The present study is focused on studying the swelling kinetics, thermal and aqueous stabilities, and determination of various forms of water in the chitosan (CS) and polyacrylonitrile (PAN) blend and semi-interpenetrating polymer network (sIPN). CS/PAN blend hydrogel films were prepared by solution casting technique. The blend film with optimum swelling properties was selected for the synthesis of sIPN. CS in the blend was crosslinked with the vapors of Glutaraldehyde (GTA) to prepare sIPN. The fabricated CS/PAN blend and sIPN hydrogels films were characterized with Fourier transform infrared (FTIR), thermal gravimetric analysis (TGA) and field emission scanning electron microscope (FESEM). The kinetics of swelling, bound and unbound waters and aqueous stability were determined experimentally. FESEM showed good miscibility between CS and PAN, FTIR showed no chemical interaction between CS and PAN; however, it did show a doublet for the sIPN, TGA showed improved thermal stability and swelling kinetic followed second order kinetics. The degree of swelling of the sIPN hydrogels samples at room temperature varied from ~2200 % (with a fair degree of stability (~30 %)) to ~1000 % (with high degree of aqueous stability (43 %)) with increase in the crosslinking time. The calculated unbound water (W_UB) max., for the blend was 52.3 % whereas for the bound (W_B) the max., was 41.9 %. However, for sIPN hydrogel films, the W_UB water decreased (max. 21.0 %) where as the W_B increased (max. 52.0 %). The decrease in W_UB and increase in the W_B is attributed to the formation of a compact structure and increase in the contact area between the water and polymers in sIPN hydrogels due to the induction of new water contacting point in these hydrogel films, respectively.     

Chemical Constituents of Lagenaria siceraria Mesocarp and Its Xanthine Oxidase and Alpha-Amylase Inhibitory Activities

The present study was conducted to identify natural products in the bioactive fractions of Lagenaria siceraria mesocarp. Column chromatography and gas chromatography-mass spectrometry (GC-MS) was used for separation and identification of phytochemicals. Methanolic extract and its fractions displayed alpha-amylase and xanthine oxidase inhibitory activities comparable to standard drugs. GC-MS analysis of the fractions and column eluates revealed the presence of a variety of chemical compounds, including 4-(methoxymethyl)phenol, 2,2?-methylenebis{6(1,1-dimethylethyl)-4-ethyl}phenol, methyl 2-hydroxy-3-phenylpropanoate, oxacyclododecane-2,8-dione, N-(2-(4-hydroxyphenyl)ethyl] acetamide, 2,3-Epoxycarane, 4-(propan-2-yl)benzaldehyde, octadec-1-ene, 4a-methyl-1-methylidene-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, 2,6-di(propan-2-yl)naphthalene, methyl 3-(4-hydroxy-3,5-dimethylphenyl)propanoate, 4,8a-dimethyl-6-(prop-1-en-2-yl)-3,5,6,7,8,8a-hexahydronaphthalen-2(1H)-one 1,3-dihydro-2-benzofuran, methyl N-hydroxybenzenecarboximidoate, linoleic acid, and palmitic acid. Many of these compounds are being reported for the first time from this plant, the presence of which may explain, at least partly, its medicinal efficacy. The fruit possesses significant anti-enzymatic properties against alpha-amylase and xanthine oxidase, and the most active compounds of mesocarp of the fruit appear in ethyl acetate fraction of its methanolic extract.     

Molecular characterization and PCR detection of a nitrogen-fixing Pseudomonas strain promoting rice growth

Nitrogen-fixing plant growth-promoting rhizobacteria (PGPR) from the genus Pseudomonas have received little attention so far. In the present study, a nitrogen-fixing phytohormone-producing bacterial isolate from kallar grass (strain K1) was identified as Pseudomonas sp. by rrs (16S ribosomal RNA gene) sequence analysis. rrs identity level was high with an uncharacterized marine bacterium (99%), Pseudomonas sp. PCP2 (98%), uncultured bacteria (98%), and Pseudomonas alcaligenes (97%). Partial nifH gene amplified from strain K1 showed 93% and 91% sequence similarities to those of Azotobacter chroococcum and Pseudomonas stutzeri, respectively. The effect of Pseudomonas strain K1 on rice varieties Super Basmati and Basmati 385 was compared with those of three non-Pseudomonas nitrogen-fixing PGPR (Azospirillum brasilense strain Wb3, Azospirillum lipoferum strain N4 and Zoogloea strain Ky1) used as single-strain inoculants. Pseudomonas sp. K1 was detected in the rhizosphere of inoculated plants by enrichment culture in nitrogen-free growth medium, which was followed by observation under the microscope as well as by PCR using a rrs-specific primer. For both rice varieties, an increase in shoot biomass and/or grain yield over that of noninoculated control plants was recorded in each inoculated treatment. The effect of Pseudomonas strain K1 on grain yield was comparable to those of A. brasilense Wb3 and Zoogloea sp. Ky1 for both rice varieties. These results show that nitrogen-fixing pseudomonads deserve attention as potential PGPR inoculants for rice.     

生物炭及生物硝化抑制剂添加对黄土区设施菜地土壤温室气体排放的影响

通过室内培养试验研究生物炭及生物硝化抑制剂添加对黄土区设施菜地土壤N2O和CO₂排放的影响,并与化学合成硝化抑制剂作对比。试验设置6个处理:不施肥(CK)、施氮(N)、施氮+生物炭(N+BC)、施     

Maize Hybrid Variability for Transpiration Decrease with Progressive Soil Drying

Drought is ubiquitous in rainfed cropping systems and often limits maize yields. The sensitivity of transpiration response early in progressive soil drying is a trait with potential to improve crop drought resistance. Simulation studies demonstrated that increased sensitivity to drying soil leading to restricted transpiration rates results in conservation of soil water during vegetative stages for possible use during grain filling. In contrast to other crops, there have been no studies characterizing genotypic variability for this trait in maize. Experiments in controlled environments were conducted to characterize the fraction of transpirable soil water (FTSW) threshold on drying soil for 36 hybrids selected for variation in the field for drought resistance, regions of adaptation and stay green. While FTSW thresholds varied among hybrids from 0.60 to 0.33, these thresholds were not uniformly associated with level of drought resistance in the field. Nevertheless, this study demonstrated a high FTSW threshold corresponded with drought resistance observed in some modern maize germplasm (hybrids #7, 17, 24, 27 and 32). This knowledge can enable breeding work seeking to exploit this adaptive trait to improved drought tolerance in low threshold FTSW germplasm.     

Molecular simulations probing Kushecarpin A as a new lipoxygenase inhibitor

Zizyphus oxyphylla Edgew is used in Pakistan as a folk medicinal remedy for inflammatory conditions, pains especially rheumatic pain, fevers, allergy and diabetes. The aim of the current study was to scientifically validate the folk use of Z. oxyphylla Edgew by using the isolated compound in vitro and in vivo levels. Kushecarpin A was isolated from ethyl acetate fraction of the plant crude extract. Molecular docking simulations predicted Kushecarpin A as a potential new lipoxygenase (LOX) inhibitor. Kushecarpin A showed significant lipoxygenase inhibition (IC50: 7.2 ± 0.02 μM) thus validated computational prediction. It also exhibited significant and highly significant inhibition (p < 0.05 and p < 0.01) of carrageenan-induced hind paw oedema at the doses of 5, 10 and 20 mg/kg. Kushecarpin A seems to be a potentially new anti-inflammatory compound responsible for anti-inflammatory activities of Z. oxyphylla Edgew. In vitro and in vivo anti-inflammatory inflammatory activities were found in good agreement with the folk medicinal use of Z. oxyphylla Edgew in inflammatory disorders.     

Fate and interaction with native soil N of ammonium N applied to wetland rice (Oryza sativa L.) grown under saline and non-saline conditions

Summary The effect of salts on the balance of fertilizer N applied as ¹⁵N-labelled ammonium sulphate and its interaction with native soil N was studied in a pot experiment using rice (Oryza sativa L.) as a test crop. The rice crop used 26%–40% of the applied N, the level of applied N and salts showing no significant bearing on the uptake of fertilizer N. Losses of fertilizer N ranged between 54% and 68% and only 5%–8% of the N was immobilized in soil organic matter. Neither the salts nor the rate of N application had any significant effect on fertilizer N immobilization. The effective use of fertilizer N (fertilizer N in grain/fertilizer N in whole plant) was, however, better in the non-saline soil. The uptake of unlabelled N (N mineralized from soil organic matter and that originating from biological N₂ fixation in thes rhizosphere) was inhibited in the presence of the salts. However, in fertilized soil, the uptake of unlabelled N was significantly enhanced, leading to increased A values [(1-% Ndff/% Ndff)x N fertilizer applied, where Ndff is N derived from fertilizer], an index of interaction with the added N. This added N interaction increased with increasing levels of added N. Since the extra unlabelled N taken up by fertilized plants was greater than the fertilizer N immobilized, and the root biomass increased with increasing levels of added N, a greater part of the added N interaction was considered to be real, any contribution by an apparent N interaction (pool substitution or isotopic displacement) to the total calculated N interaction being fairly small. Under saline conditions, for the same level of fertilizer N addition, the added N interaction was lower, and this was attributed to a lower level of microbial activity, including mineralization of native soil N, rootdriven immobilization of applied N, and N₂ fixation.     

Growth,survival, and heavy metal (Cd and Ni) uptake of spinach (Spinacia oleracea) and fenugreek (Trigonella corniculata) in a biochar‐amended sewage‐irrigated contaminated soil

Irrigation of arable land with contaminated sewage waters leads to the accumulation of trace metals in soils with subsequent phyto‐/zootoxic consequences. In this study, biochar derived from cotton sticks was used to amend an agricultural silt‐loam soil that had been previously irrigated with trace metal contaminated sewage waters. Metal accumulation and toxicity to spinach (Spinacia oleracea) and fenugreek (Trigonella corniculata) was investigated by measuring concentrations of Cd and Ni in plant tissues and various photosynthetic and biochemical activities of plants. Positive impacts of biochar on both spinach and fenugreek were observed in terms of biomass production that increased from 29% to 36% in case of spinach, while for fenugreek this increase was 32% to 36%. In the control treatment there was an increase in malondialdihyde, soluble sugar, and ascorbic acid contents, indicating heavy metal stress. Biochar applications increased soluble proteins and amino acids in plants and reduced the uptake of Cd from 5.42 mg kg^?1 at control to 3.45 mg kg^?1 at 5% biochar amended soil and Ni (13.8 mg kg^?1 to 7.3 mg kg^?1 at 5% biochar) by the spinach plants. In fenugreek, the Cd was reduced from 7.72 mg kg^?1 to 3.88 mg kg^?1 and reduction in Ni was from 15.45 mg kg^?1 to 9.46 mg kg^?1 at 5% biochar treated soil, reducing the possibility of transfer up the food chain. This study demonstrates that the use of biochar made from cotton‐sticks, as an amendment to arable soils that have received contaminated irrigation water, could improve plant growth and decrease Cd and Ni uptake to crops, alleviating some of the negative impacts of using sewage waters on arable land.     

Regeneration in sugarcane via somatic embryogenesis and genomic instability in regenerated plants

In the present study, embryogenic calli of sugarcane variety BL4 were induced using 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin in different concentrations and combinations. In contrast to earlier studies, embryogenic callus sectors were identified and isolated microscopically within 1–2 weeks. Subsequently, 51 media formulations were used for regeneration of proliferated embryogenic callus, using MS medium supplemented with three different cytokinins [kinetin, 6-Benzylamino purine (BAP), and thidiazuron (TDZ)] and auxins (IAA/NAA and IBA) in different combination and concentrations. After acclimatization, the genomic DNA of regenerated plants was studied to explore the insertion polymorphism of transposable elements in order to ascertain the variation among somaclones. Though low concentration of kinetin with 2,4-D was found supportive to embryogenic callus development, the highest number of regenerated plantlets was observed using BAP (1 μM), however the plantlets had very low fresh weight (2.2 g). Conversely, TDZ alone supported a significant increase in the number of plantlets regenerated (38–40) with higher fresh weight. The somaclones generated during this study showed considerable positional polymorphism of activator-like transposable elements possibly due to the stress associated with in vitro culture. This study provides a procedure to produce regenerated sugarcane plants from embryogenic callus in a relatively short time.