Impact of Different Rates of Nitrogen Supplementation on Soil PhysicochemicalProperties and Microbial Diversity in Goji Berry

Goji berry(Lycium barbarum L.)is substantially dependent on nitrogen fertilizer application,which can signifi-cantly enhance fruit yield and Goji berry industrial development in Ningxia,China.This study aimed to analyze the functions of differential nitrogen application rates including low(N1),medium(N2),and high(N3)levels in soil microbial community structure(bacterial and fungal)at 2 diverse soil depths(0-20,20-40 cm)through high-throughput sequencing technology by targeting 16S RNA gene and ITS1&ITS2 regions.All the observed physicochemical parameters exhibited significant improvement(p<0.05)with increased levels of nitrogen and the highest values for most parameters were observed at N2.However,pH decreased(p<0.05)gradually.The alpha and beta diversity analyses for bacterial and fungal communities’metagenome displayed more similarities than differences among all groups.The top bacterial and fungal phyla and genera suggested no obvious(p>0.05)differences among three group treatments(N1,N2,and N3).Furthermore,the functional enrichment analysis demonstrated significant(p<0.05)enrichment of quorum sensing,cysteine and methionine metabolism,and transcriptional machinery for bacterial communities,while various saprotrophic functional roles for fungal communities.Conclusively,moderately reducing the use of N-supplemented fertilizers is conducive to increasing soil nitrogen utilization rate,which can contribute to sustainable agriculture practices through improved soil quality,and microbial community structure and functions.

ERF5.1 modulates carotenoid accumulation by interacting with CCD4.1 in Lycium

Carotenoids are important natural pigments and have medical and health functions for humans.Carotenoid cleavage dioxygenase 4(CCD4)and ethylene responsive factor(ERF)participate in carotenoid metabolism,but their roles in Lycium have not been discovered.Here,we annotated LbCCDs from the Lycium reference genome and found that LbCCD4.1 expression was significantly correlated with the carotenoid metabolites during Lycium five fruit developmental stages.Over-expression of LbCCD4.1 in NQ’s leaves resulted in a series of significantly lower contents of carotenoid metabolites,includingβ-carotene andβ-cryptoxanthin.Moreover,LbERF5.1,a transcription factor belonging to the ERF family that was located in the nucleus,was isolated.Significant reductions in the carotenoids,especially lutein,violaxanthin and their derivatives,were observed in over-expressing ERF5.1 transgenic NQ’s leaves.Over-expression or virus-induced gene silencing of LbERF5.1 in NQ’s leaves induced a consistent up-or down-expression,respectively,of LbCCD4.1.Furthermore,yeast one-hybrid and dual-luciferase reporter assays showed that ERF5.1 interacted with the promoter of CCD4.1 to increase its expression,and LbERF5.1 could bind to any one of the three predicted binding sites in the promoter of LbCCD4.1.A transcriptome analysis of LbERF5.1 and LbCCD4.1 over-expressed lines showed similar global transcript expression,and geranylgeranyl diphosphate synthase,phytoene synthase,lycopeneδ-cyclase cytochrome,cytochrome P450-type monooxygenase 97A,cytochrome P450-type monooxygenase 97C,and zeaxanthin epoxidase in the carotenoid biosynthesis pathway were differentially expressed.In summary,we uncovered a novel molecular mechanism of carotenoid accumulation that involved an interaction between ERF5.1 and CCD4.1,which may be used to enhance carotenoid in Lycium.

Microwave-assisted preparation and in vitro characterizations of doped superparamagnetic ferrite nanoclusters

In order to obtain nanomaterials with superparamagnetism and high saturation magnetization, Mn-doped or Zn-doped superparamagnetic ferrite nanoclusters(Mn-FNs or Zn-FNs) were prepared by microwave-assisted solvothermal method in this study. Preliminary investigations were performed by transmission electron microscopy(TEM) and dynamic light scattering(DLS) instrument to observe the morphology and measure the particle size, respectively. Afterwards, Zn-FNs were chosen to be further characterized in vitro due to their better morphology and dispersity than Mn-FNs. The subsequent characterizations included crystalline phase, metal content and magnetic properties by X-ray diffractometer(XRD), inductively coupled plasma-mass spectrometry(ICP-MS) and vibrating sample magnetometer(VSM), respectively. The results showed that Zn-FNs had a cluster-like structure assembled by multiple nanoparticles. Zn-FNs were spherical in shape with good dispersity and relatively uniform particle size. Zn was successfully doped in Zn-FNs which demonstrated spinel structure and excellent magnetic properties. Therefore, Zn-FNs had a favorable application prospect as a new type of magnetic nanomaterial.

A simple and sensitive gradient elution liquid chromatography with tandem mass spectrometry(LC-MS/MS) method for the quantification of doxorubicin in rabbit plasma

In the present study, we developed and validated a simple and sensitive gradient elution liquid chromatography tandem mass spectrometry （LC-MS/MS） method for quantification of doxorubicin in rabbit plasma. Daunorubicin was used as an internal standard （IS）. The doxorubicin and IS were extracted with ethyl acetate from plasma samples. The chromatographic separations were achieved on a C18 column （2.1 mm×50 mm, 2.5μm） configured with a C18 guard column （2.1 mm×10 mm, 2.5 μm）. The mobile phase of 0.1% formic acid-water solution and acetonitrile was delivered using a gradient elution program at a flow rate of 0.4 mL/min. The temperature for column was maintained at 40 ℃. The electrospray ionization （ESI） source was operated in the positive ion mode, and the quantification was conducted using multiple reaction monitoring （MRM） of the transitions m/z 544.07→396.96 and m/z 528.06→321.05 for doxorubicin and IS, respectively. The calibration curve of doxorubicin was linear （r 〉 0.999） within the range of 2-600 ng/mL. The lower limit of quantification was 2 ng/mL. The relative errors of intra-day and inter-day accuracies ranged from -2.48% to 0.18% and from -3.78% to 1.94%, respectively. The relative standard deviations of intra-day and inter-day precisions were less than 8.65% and 6.41%, respectively. The method exhibited satisfactory results in terms of specificity, sensitivity, matrix effect, recovery and stability. The newly developed LC-MS/MS method was reliable to monitor doxorubicin concentrations in rabbit plasma.