The combination of a microbial and a non-microbial biostimulant increases yield in lettuce(Lactuca sativa) under salt stress conditions by up-regulating cytokinin biosynthesis

Salinization poses a significant challenge in agriculture, exacerbated by anthropogenic global warming. Biostimulants, derived from living microorganisms or natural extracts, have emerged as valuable tools for conventional and organic agriculture.However, our understanding of the molecular mechanisms underlying the effects of biostimulants is very limited, especially in crops under real cultivation conditions. In this study, we adopted an integrative approach to investigate the effectiveness of the combined application of plant growth-promoting bacterium(Bacillus megaterium strain BM08) and a non-microbial biostimulant under control conditions(normal watering) and salt stress. After confirming the yield increase under both conditions, we investigated the molecular mechanisms underlying the observed effect by measuring a number of physiological parameters(i.e., lipid peroxidation, antioxidants,chlorophylls, total phenolics and phytohormone content), as well as RNA sequencing and primary metabolite analyses. Our findings reveal that the combined effect of the microbial and non-microbial biostimulants led to a decrease in the antioxidant response and an up-regulation of genes involved in cytokinin biosynthesis under salt stress conditions.This, in turn, resulted in a higher concentration of the bioactive cytokinin, isopentenyladenosine, in roots and leaves and an increase in γ-aminobutyric acid, a non-proteic amino acid related to abiotic stress responses. In addition, we observed a decrease in malic acid, along with an abscisic acid(ABA)-independent up-regulation of SR-kinases, a family of protein kinases associated with abiotic stress responses. Furthermore, we observed that the single application of the non-microbial biostimulant triggers an ABA-dependent response under salt stress;however, when combined with the microbial biostimulant, it potentiated the mechanisms triggered by the BM08 bacterial strain. This comprehensive investigation shows that the combination of two biostimulants is able to elicit a cytokinin-dependent response that may explain the observed yield increase under salt stress conditions.

Fractionation and characterization of bio-oil from microwave-assisted pyrolysis of corn stover

Bio-oil from thermochemical conversion of biomass is a complex mixture of polar and non-polar compounds,and cannot be used directly as a combustion engine fuel due to its low heating value,high viscosity,chemical instability,and incomplete volatility.Mixed-solvent extraction was developed to fractionate bio-oils from microwave-assisted pyrolysis of corn stover to produce light oil(a mixture of light and inflammable fuel oil components from bio-oil)with low viscosity and low combustion residue,and high value chemicals.Different fractions from bio-oil were characterized using GC/MS and TG,and a major chemical(hydroxy-butanedioic acid diethyl ester)was separated.

CRISPR-Cas9 sgRNA design and outcome assessment: Bioinformatics tools and aquaculture applications

The CRISPR-Cas9 genome editing has many advantages over its counterparts and could ultimately assist in disease control and molecular breeding in aquaculture.Single-guide RNA(sgRNA)design is presumably the most crucial task in a typical CRISPR-Cas9 experiment;an understanding of algorithms behind sgRNA design programs is thus essential for improved efficacy and specificity.We focus this review on the bioinformatics aspects in genome editing experiments and describe commonly used computational approaches and tools of sgRNA design and outcome assessment.We show an example of sgRNA design with optimal parameter settings and appropriate interpretation of results and present a brief overview of CRISPR-Cas9 applications,such as genetic improvement and sustainability in aquaculture.We discuss challenging issues,particularly in the context of computational biology,in the use of computational tools in CRISPR-based genome editing.This review provides a synthesis of bioinformatics tools used for CRISPR-Cas9 sgRNA design and outcome assessment and offers a general view of CRISPR-based applications in farmed fish,which are expected to facilitate genome editing programs and hence improve aquaculture breeding,production,and sustainability.

Integrative multi-platform meta-analysis of hepatocellular carcinoma gene expression profiles for identifying prognostic and diagnostic biomarkers

Hepatocellular carcinoma(HCC),as one of the most com-mon types of primary liver cancer(PLV),accounts for approximately 75%e90%of all PLV cases identified worldwide.^(1)Currently,the most effective treatment for HCC patients constitutes liver transplantation.However,due to high rates of recurrence a poor prognosis is predicted.^(2)Therefore,accurate HCC biomarkers are ur-gently needed to develop innovative therapeutics.Most of earlier investigations to identify biomarkers have been severely limited by sample size.