Endogenous biosynthesis of docosahexaenoic acid(DHA)regulates fish oocyte maturation by promoting pregnenolone production

Omega-3 polyunsaturated fatty acids(n-3 PUFAs),particularly docosahexaenoic acid(22:6n-3,DHA),play crucial roles in the reproductive health of vertebrates,including humans.Nevertheless,the underlying mechanism related to this phenomenon remains largely unknown.In this study,we employed two zebrafish genetic models,i.e.,elovl2^(-/-)mutant as an endogenous DHAdeficient model and fat1(omega-3 desaturase encoding gene)transgenic zebrafish as an endogenous DHA-rich model,to investigate the effects of DHA on oocyte maturation and quality.Results show that the elovl2^(-/-)mutants had much lower fecundity and poorer oocyte quality than the wild-type controls,while the fat1 zebrafish had higher fecundity and better oocyte quality than wildtype controls.DHA deficiency in elovl2^(-/-)embryos led to defects in egg activation,poor microtubule stability,and reduced pregnenolone levels.Further study revealed that DHA promoted pregnenolone synthesis by enhancing transcription of cyp11a1,which encodes the cholesterol side-chain cleavage enzyme,thereby stabilizing microtubule assembly during oogenesis.In turn,the hypothalamic-pituitary-gonadal axis was enhanced by DHA.In conclusion,using two unique genetic models,our findings demonstrate that endogenously synthesized DHA promotes oocyte maturation and quality by promoting pregnenolone production via transcriptional regulation of cyp11a1.

Knock-out of GhPDCT with the CRISPR/Cas9 system increases the oleic acid content in cottonseed oil

Cotton is a pivotal economic crop for natural textile fibers that also serves as an important source of edible oil(Long et al.2023).Cottonseed oil contains approximately14%oleic acid and 59%linoleic acid.An increase in monounsaturated fatty acids,particularly oleic acid,enhances the oxidative stability and nutritional value of edible oil(Chen et al.2021).

Epigenetic control on transcription of vernalization genes and whole-genome gene expression profile induced by vernalization in common wheat

Vernalization is necessary for winter wheat to flower.However,it is unclear whether vernalization is also required for spring wheat,which is frequently sown in fall,and what molecular mechanisms underlie the vernalization response in wheat varieties.In this study,we examined the molecular mechanisms that regulate vernalization response in winter and spring wheat varieties.For this purpose,we determined how major vernalization genes(VRN1,VRN2,and VRN3)respond to vernalization in these varieties and whether modifications to histones play a role in changes in gene expression.We also identified genes that are differentially regulated in response to vernalization in winter and spring wheat varieties.We found that in winter wheat,but not in spring wheat,VRN1 expression decreases when returned to warm temperature following vernalization.This finding may be associated with differences between spring and winter wheat in the levels of tri-methylation of lysine 27 on histone H3(H3K27me3)and tri-methylation of lysine 4 on histone H3(H3K4me3)at the VRN1 gene.Analysis of winter wheat transcriptomes before and after vernalization revealed that vernalization influences the expression of several genes,including those involved in leucine catabolism,cysteine biosynthesis,and flavonoid biosynthesis.These findings provide new candidates for further study on the mechanism of vernalization regulation in wheat.

Heat-inducible SlWRKY3 confers thermotolerance by activating the SlGRXS1 gene cluster in tomato

High temperature stress is one of the major environmental factors that affect the growth and development of plants. Although WRKY transcription factors play a critical role in stress responses, there are few studies on the regulation of heat stress by WRKY transcription factors,especially in tomato. Here, we identified a group I WRKY transcription factor, SlWRKY3, involved in thermotolerance in tomato. First, SlWRKY3 was induced and upregulated under heat stress. Accordingly, overexpression of SlWRKY3 led to an increase, whereas knock-out of SlWRKY3 resulted in decreased tolerance to heat stress. Overexpression of SlWRKY3 accumulated less reactive oxygen species(ROS), whereas knock-out of SlWRKY3 accumulated more ROS under heat stress. This indicated that SlWRKY3 positively regulates heat stress in tomato. In addition,SlWRKY3 activated the expression of a range of abiotic stress-responsive genes involved in ROS scavenging, such as a SlGRXS1 gene cluster.Further analysis showed that SlWRKY3 can bind to the promoters of the SlGRXS1 gene cluster and activate their expression. Collectively, these results imply that SlWRKY3 is a positive regulator of thermotolerance through direct binding to the promoters of the SlGRXS1 gene cluster and activating their expression and ROS scavenging.

Identification of new cotton fiber-quality QTL by multiple genomic analyses and development of markers for genomic breeding

Cotton fiber is one of the main raw materials for the textile industry.In recent years,many cotton fiber quality QTL have been identified,but few were applied in breeding.In this study,a genome wide association study(GWAS)of fiber-quality traits in 265 upland cotton breeding intermediate lines(GhBreeding),combined with genome-wide selective sweep analysis(GSSA)and genomic selection(GS),revealed 25 QTL.Most of these QTL were ignored by only using GWAS.The CRISPR/Cas9 mutants of GhMYB_D13 had shorter fiber,which indicates the credibility of QTL to a certain extent.Then these QTL were verified in other cotton natural populations,5 stable QTL were found having broad potential for application in breeding.Additionally,among these 5 stable QTL,superior genotypes of 4 showed an enrichment in most improved new varieties widely cultivated currently.These findings provide insights for how to identify more QTL through combined multiple genomic analysis to apply in breeding.

Anti-infective immune functions of type Ⅳ interferon in grass carp(Ctenopharyngodon idella):A novel antibacterial and antiviral interferon in lower vertebrates

Type Ⅳ interferon(IFN-υ)is a recently discovered cytokine crucial for host defense against viral infections.However,the role and mechanisms of IFN-υin bacterial infections remain unexplored.This study investigated the antibacterial and antiviral functions and mechanisms of grass carp(Ctenopharyngodon idella)IFN-υ(CiIFN-υ)both in vivo and in vitro.The CiIFN-υgene was first identified and characterized in grass carp.Subsequently,the immune expression of CiIFN-υsignificantly increased following bacterial challenge,indicating its response to bacterial infections.The eukaryotic recombinant expression plasmid of CiIFN-υwas then constructed and transfected into fathead minnow(FHM)cells.Supernatants were collected and incubated with four bacterial strains,followed by plate spreading and colony counting.Results indicated that CiIFN-υexhibited more potent antibacterial activity against gram-negative bacteria compared to gram-positive bacteria and aggregated gram-negative bacteria but not gram-positive bacteria.In vivo experiments further confirmed the antibacterial function,showing high survival rates,low tissue edema and damage,reduced tissue bacterial load,and elevated proinflammatory response at the early stages of bacterial infection.In addition,the antiviral function of CiIFN-υwas confirmed through in vitro and in vivo experiments,including crystal violet staining,survival rates,tissue viral burden,and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR).This study highlights the antibacterial function and preliminary mechanism of IFN-υ,demonstrating that IFN-υpossesses dual functions against bacterial and viral infections.

Comprehensive integration of single-cell transcriptomic data illuminates the regulatory network architecture of plant cell fate specification

Plant morphogenesis relies on precise gene expression programs at the proper time and position which is orchestrated by transcription factors(TFs)in intricate regulatory networks in a cell-type specific manner.Here we introduced a comprehensive single-cell transcriptomic atlas of Arabidopsis seedlings.This atlas is the result of meticulous integration of 63 previously published scRNA-seq datasets,addressing batch effects and conserving biological variance.This integration spans a broad spectrum of tissues,including both below-and above-ground parts.Utilizing a rigorous approach for cell type annotation,we identified 47 distinct cell types or states,largely expanding our current view of plant cell compositions.We systematically constructed cell-type specific gene regulatory networks and uncovered key regulators that act in a coordinated manner to control cell-type specific gene expression.Taken together,our study not only offers extensive plant cell atlas exploration that serves as a valuable resource,but also provides molecular insights into gene-regulatory programs that varies from different cell types.

Utilizing auxin dwarf genes to optimize seed yield and lodging resistance in rapeseed

Direct-seeding rapeseed production at high plant density raises the risk of lodging.We investigated the use of dwarf genes to improve rapeseed plant architecture to balance yield and lodging.Three genotypes with different plant architectures(dwarf sca^(HS5),semi-dwarf+/sca^(HS5),and tall ^(HS5))were evaluated under varying nitrogen rates(N1,N2,and N3:120,240,and 360 kg N ha^(-1))and plant densities(D1,D2,and D3:15,45,and 75 plants m^(-2))from 2019 to 2022.The results showed that increasing N rate positively influenced yield while decreasing lodging resistance in all genotypes.Increasing plant density(D2-D3)enhanced lodging resistance and yield in sca^(HS5) and+/sca^(HS5),but reduced yield in ^(HS5).Compared to the two parents,+/sca^(HS5) exhibited moderate expressions of IAA3,GH3.15,and SAUR30 in stems under N2D3,resulting in reduced plant height and increased compactness.Additionally,+/sca^(HS5) had a thicker silique layer than ^(HS5) by 14.7%,and it had a significant correlation between branch height/angle and yield.Increasing N rate led to increased lignin and pectin contents,while cellulose content decreased.Increasing plant density resulted in greater stem cellulose content and CSLA3/7 expression in sca^(HS5) and+/sca^(HS5),but decreased in ^(HS5).Compared to ^(HS5),+/sca^(HS5) exhibited higher expressions of ARAD1 and GAUT4,along with a 51.1%increase in pectin content,leading to improved lodging resistance under N2D3.Consequently,+/sca^(HS5) showed a 46.4%higher yield and 38.9%lodging resistance than ^(HS5) under N2D3,while sca^(HS5) demonstrated strong lodging resistance but lower yield potential.Overall,this study underscores the potential of utilizing auxin dwarf genes to optimize the trade-off between yield and lodging resistance in rapeseed and the possibility of maximizing yield potential by optimizing the plant architecture of+/sca^(HS5) through nitrogen reduction and dense planting.

Apical meristem transcriptome analysis identifies a role for the blue light receptor gene GhFKF1 in cotton architecture development

Cotton architecture is determined by the differentiation fate transition of axillary meristem(AM),and influences cotton yield and the efficiency of mechanized harvesting.We observed that the initiation of flowering primordium was earlier in early-maturing than that in late-maturing cultivars during the differentiation and development of AM.The RNA-Seq and expression level analyses showed that genes FLAVIN BINDING,KELCH REPEAT,F-BOX1(GhFKF1),and GIGANTEA(GhGI)were in response to circadian rhythms,and involved in the regulation of cotton flowering.The gene structure,predicted protein structure,and motif content analyses showed that in Arabidopsis,cotton,rapseed,and soybean,proteins GhFKF1 and GhGI were functionally conserved and share evolutionary origins.Compared to the wild type,in GhFKF1 mutants that were created by the CRISPR/Cas9 system,the initiation of branch primordium was inhibited.Conversely,the knocking out of GhGI increased the number of AM differentiating into flower primordium,and there were much more lateral branch differentiation and development.Besides,we investigated that proteins GhFKF1 and GhGI can interact with each other.These results suggest that GhFKF1 and GhGI are key regulators of cotton architecture development,and may collaborate to regulate the differentiation fate transition of AM,ultimately influencing plant architecture.We describe a strategy for using the CRISPR/Cas9 system to increase cotton adaptation and productivity by optimizing plant architecture.

A novel histone methyltransferase gene Cg SDG40 positively regulates carotenoid biosynthesis during citrus fruit ripening

The flesh color of pummelo(Citrus maxima)fruits is highly diverse and largely depends on the level of carotenoids,which are beneficial to human health.It is vital to investigate the regulatory network of carotenoid biosynthesis to improve the carotenoid content in pummelo.However,the molecular mechanism underlying carotenoid accumulation in pummelo is not fully understood.In this study,we identified a novel histone methyltransferase gene,CgSDG40,involved in carotenoid regulation by analyzing the flesh transcriptome of typical white-fleshed pummelo,red-fleshed pummelo and extreme-colored F1 hybrids from a segregated pummelo population.Expression of CgSDG40 corresponded to flesh color change and was highly coexpressed with CgPSY1.Interestingly,CgSDG40 and CgPSY1 are located physically adjacent to each other on the chromosome in opposite directions,sharing a partially overlapping promoter region.Subcellular localization analysis indicated that CgSDG40 localizes to the nucleus.Overexpression of CgSDG40 significantly increased the total carotenoid content in citrus calli relative to that in wild type.In addition,expression of CgPSY1 was significantly activated in overexpression lines relative to wild type.Taken together,our findings reveal a novel histone methyltransferase regulator,CgSDG40,involved in the regulation of carotenoid biosynthesis in citrus and provide new strategies for molecular design breeding and genetic improvement of fruit color and nutritional quality.

foxl2l is a germ cell-intrinsic gatekeeper of oogenesis in zebrafish

Zebrafish serve as a valuable model organism for studying germ cell biology and reproductive processes.The AB strain of zebrafish is proposed to exhibit a polygenic sex determination system,where most males initially develop juvenile ovaries before committing to male fate.In species with chromosomal sex determination,gonadal somatic cells are recognized as key determinants of germ cell fate.Notably,the loss of germ cells in zebrafish leads to masculinization,implying that germ cells harbor an intrinsic feminization signal.However,the specific signal triggering oogenesis in zebrafish remains unclear.In the present study,we identified foxl2l as an oocyte progenitor-specific gene essential for initiating oogenesis in germ cells.Results showed that foxl2l-knockout zebrafish bypassed the juvenile ovary stage and exclusively developed into fertile males.Further analysis revealed that loss of foxl2l hindered the initiation of oocyte-specific meiosis and prevented entry into oogenesis,leading to premature spermatogenesis during early gonadal development.Furthermore,while mutation of the pro-male gene dmrt1 led to fertile female differentiation,simultaneous disruption of foxl2l in dmrt1 mutants completely blocked oogenesis,with a large proportion of germ cells arrested as germline stem cells,highlighting the crucial role of foxl2l in oogenesis.Overall,this study highlights the unique function of foxl2l as a germ cell-intrinsic gatekeeper of oogenesis in zebrafish.

Combining genome-wide association study and transcriptome analysis to identify molecular markers and genetic basis of population-asynchronous ovarian development in Coilia nasus

Coilia nasus,a migratory fish species found in the middle and lower reaches of the Yangtze River and along offshore areas of China,possesses considerable aquacultural and economic potential.However,the species faces challenges due to significant variation in the gonadal development rate among females,resulting in inconsistent ovarian maturation times at the population level,an extended reproductive period,and limitations on fish growth rate due to ovarian prematurity.In the present study,we combined genome-wide association study(GWAS)and comparative transcriptome analysis to investigate the potential single nucleotide polymorphisms(SNPs)and candidate genes associated with population-asynchronous ovarian development in C.nasus.Genotyping of the female population based on whole-genome resequencing yielded 2120695 high-quality SNPs,39 of which were suggestively associated with ovarian development.Of note,a significant SNP peak on LG21 containing 30 suggestively associated SNPs was identified,with cpne5a determined as the causal gene of the peak.Therefore,single-marker and haplotype association analyses were performed on cpne5a,revealing four genetic markers(P<0.05)and seven haplotypes(r2>0.9)significantly associated with the phenotype.Comparative transcriptome analysis of precociously and normally maturing individuals screened out 29 and 426 overlapping differentially expressed genes in the brain and ovary,respectively,between individuals of different body sizes.Integrating the GWAS and transcriptome analysis results,this study identified genes and pathways related to hypothalamic-pituitary-gonadal axis hormone secretion,extracellular matrix,angiogenesis,and gap junctions involved in population-asynchronous ovarian development.The insights gained from this study provide a basis for a deeper understanding of the molecular mechanisms underlying ovarian development in fish and may facilitate the genetic breeding of C.nasus strains exhibiting population-synchronous ovarian development in the future.

Characterization of a 4.1 Mb inversion harboring the stripe rust resistance gene YR86 on wheat chromosome 2AL

Wheat cultivar Zhongmai 895 was earlier found to carry YR86 in an 11.6 Mb recombination-suppressed region on chromosome 2AL when crossed with Yangmai 16.To fine-map the YR86 locus,we developed two large F2 populations from crosses Emai 580/Zhongmai 895 and Avocet S/Zhongmai 895.Remarkably,both populations exhibited suppressed recombination in the same 2AL region.Collinearity analysis across Chinese Spring,Aikang 58,and 10+wheat genomes revealed a 4.1 Mb chromosomal inversion spanning 708.5-712.6 Mb in the Chinese Spring reference genome.Molecular markers were developed in the breakpoint and were used to assess a wheat cultivar panel,revealing that Chinese Spring,Zhongmai 895,and Jimai 22 shared a common sequence named InvCS,whereas Aikang 58,Yangmai 16,Emai 580,and Avocet S shared the sequence named InvAK58.The inverted configuration explained the suppressed recombination observed in all three bi-parental populations.Normal recombination was observed in a Jimai 22/Zhongmai 895 F2 population,facilitating mapping of YR86 to a genetic interval of 0.15 cM corresponding to 710.27-712.56 Mb falling within the inverted region.Thirty-three high-confidence genes were annotated in the interval using the Chinese Spring reference genome,with six identified as potential candidates for YR86 based on genome and transcriptome analyses.These results will accelerate map-based cloning of YR86 and its deployment in wheat breeding.

Utilizing resequencing big data to facilitate Brassica vegetable breeding:tracing introgression pedigree and developing highly specific markers for clubroot resistance

Clubroot caused by Plasmodiophora brassicae is a devastating disease of Cruciferous crops.Developing cultivars with clubroot resistance(CR)is the most effective control measure.For the two major Brassica vegetable species B.rapa and B.oleracea,several commercial cultivars with unclear CR pedigrees have been intensively used as CR donors in breeding.However,the continuous occurrence of CR-breaking makes the CR pedigree underlying these cultivars one of the breeders'most urgent concerns.The complex intraspecific diversity of these two major Brassica vegetables has also limited the applicability of CR markers in different breeding programs.Here we first traced the pedigree underlying two kinds of CR that have been widely applied in breeding by linkage and introgression analyses based on public resequencing data.In B.rapa,a major locus CRzi8 underlying the CR of the commercial CR donor‘DegaoCR117’was identified.CRzi8 was further shown to have been introgressed from turnip(B.rapa ssp.rapifera)and that it carried a potential functional allele of Crr1a.The turnip introgression carried CRb^(c),sharing the same coding sequence with the CRb that was also identified from chromosome C07 of B.oleracea CR cultivars with different morphotypes.Within natural populations,variation analysis of linkage intervals of CRzi8,PbBa8.1,CRb,and CRb^(c)yielded easily resolved InDel markers(>20 bp)for these fundamental CR genes.The specificity of these markers was tested in diverse cultivars panels,and each exhibited high reliability in breeding.Our research demonstrates the value of the practice of applying resequencing big data to solve urgent concerns in breeding programs.

Ferroptosis is involved in deoxynivalenol-induced intestinal damage in pigs

Background Deoxynivalenol(DON)is a widespread issue for feed and food safety,leading to animal and human health risks.The objective of this study was to determine whether ferroptosis is involved in DON-induced intestinal injury in piglets.Three groups of 21-day-old male weanling piglets(n 4,serum and small intestines were=7/group)were fed a control diet,or diet adding 1.0 or 3.0 mg DON/kg.At week collected to assay for biochemistry,histology,redox status and ferroptosis-related genes expression.In addition,the involvement of ferroptosis and the role of FTL gene in DON-induced cell death were further verified in the IPEC-J2 cells.Results Compared to the control,dietary supplementation of DON at 1.0 and 3.0 mg/kg induced different degrees of damage in the duodenum,jejunum and ileum,and increased(P<0.05)serum lipopolysaccharide concentration by 46.2%-51.4%.Dietary DON supplementation at 1.0 and(or)3.0 mg/kg increased(P<0.05)concentrations of malondialdehyde(17.4%-86.5%)and protein carbonyl by 33.1%-92.3%in the duodenum,jejunum and ileum.In addition,dietary supplemented with DON upregulated(P<0.05)ferroptotic gene(DMT1)and anti-ferroptotic genes(FTL and FTH1),while downregulated(P<0.05)anti-ferroptotic genes(FPN,FSP1 and CISD1)in the duodenum of the porcine.Furthermore,the in vitro study has demonstrated that deferiprone,a potent ferroptotic inhibitor,mitigated(P<0.05)DON-induced cytotoxicity in porcine small intestinal IPEC-J2 cells.Additionally,deferiprone prevented or alleviated(P<0.05)the dysregulation of ferroptosis-related genes(ACSL4 and FTL)by DON in IPEC-J2 cells.Moreover,specific siRNA knockdown FTL gene expression compromised the DON-induced cell death in IPEC-J2 cells.Conclusions In conclusion,this study revealed that ferroptosis is involved in DON-induced intestinal damage in porcine,and sheds a new light on the toxicity of DON to piglets.

Grain yield and nitrogen use efficiency of an ultrashort-duration variety grown under different nitrogen and seeding rates in direct-seeded and double-season rice in Central China

Nitrogen(N) and seeding rates are important factors affecting grain yield and N use efficiency(NUE) in directseeded rice. However, these factors have not been adequately investigated on direct-seeded and double-season rice(DDR) in Central China. The objective of this study was to evaluate the effects of various N and seeding rates on the grain yield and NUE of an ultrashort-duration variety grown under DDR. Field experiments were conducted in 2018 in Wuxue County and 2019 in Qichun County, Hubei Province, China with four N rates and three seeding rates.The results showed that the grain yield of the ultrashort-duration variety ranged from 6.32 to 8.23 t ha–1with a total growth duration of 85 to 97 days across all treatments with N application. Grain yield was increased significantly by N application in most cases, but seeding rate had an inconsistent effect on grain yield. Furthermore, the response of grain yield to the N rates was much higher than the response to seeding rates. The moderate N rates of 100–150 and 70–120 kg N ha–1in the early and late seasons, respectively, could fully express the yield potential of the ultrashort-duration variety grown under DDR. Remarkably higher N responses and agronomic NUE levels were achieved in the early-season rice compared with the late-season rice due to the difference in indigenous soil N supply capacity(INS) between the two seasons. Seasonal differences in INS and N response should be considered when crop management practices are optimized for achieving high grain yield and NUE in ultrashort-duration variety grown under DDR.

Border effects of the main and ratoon crops in the rice ratooning system

The border effect(BE)is widely observed in crop field experiments,and it has been extensively studied in many crops.However,only limited attention has been paid to the BE of ratoon rice.We conducted field experiments on ratoon rice in Qichun County,Hubei Province,Central China in 2018 and 2019 to compare the BE in the main and ratoon crops,and to quantify the contribution of BE in the main crop to that in the ratoon crop.The BE of two hybrid varieties was measured for the outermost,second outermost,and third outermost rows in each plot of both crops.To determine the contribution of BE between the two crops,portions of hills in the outermost and second outermost rows were uprooted during the harvest of the main crop so that the second and third outermost rows then became the outermost rows in the ratoon crop.Overall,the BE on grain yield was greater in the main crop than in the ratoon crop.In the main crop,the BE on grain yield was 98.3%in the outermost row,which was explained by the BE on panicles m^(–2),spikelets/panicle,spikelets m^(–2),and total dry weight.In the ratoon crop,the BE on grain yield was reduced to 60.9 and 27.6%with and without the contribution of the BE in the main crop,respectively.Consequently,55.1%of the BE on grain yield in the ratoon crop was contributed from the main crop.High stubble dry weight and non-structural carbohydrate(NSC)accumulation at the harvest of the main crop were responsible for the contribution of BE in the main crop to that in the ratoon crop.Our results suggest that increases in stubble dry weight and NSC accumulation at the harvest of the main crop could be important strategies for developing high-yielding cropping practices in the rice ratooning system.

An efficient transient gene expression system for protein subcellular localization assay and genome editing in citrus protoplasts

Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.

Depletion of stearoyl-CoA desaturase(scd) leads to fatty liver disease and defective mating behavior in zebrafish

Stearyl coenzyme A desaturase(SCD), also known as delta-9 desaturase, catalyzes the rate-limiting step in the formation of monounsaturated fatty acids.In mammals, depletion or inhibition of SCD activity generally leads to a decrease in triglycerides and cholesteryl esters. However, the endogenous role of scd in teleost fish remains unknown. Here, we generated a zebrafish scd mutant(scd-/-) to elucidate the role of scd in lipid metabolism and sexual development. Gas chromatography-mass spectrometry(GC-MS) showed that the scd-/- mutants had increased levels of saturated fatty acids C16:0 and C18:0, and decreased levels of monounsaturated fatty acids C16:1 and C18:1. The mutant fish displayed a short stature and an enlarged abdomen during development. Unlike Scd-/ -mammals, the scd-/- zebrafish showed significantly increased fat accumulation in the whole body,especially in the liver, leading to hepatic mitochondrial dysfunction and severe cell apoptosis.Mechanistically, srebf1, a gene encoding a transcriptional activator related to adipogenesis,acc1 and acaca, genes involved in fatty acid synthesis, and dgat2, a key gene involved in triglyceride synthesis, were significantly upregulated in mutant livers to activate fatty acid biosynthesis and adipogenesis. The scd-/- males exhibited defective natural mating behavior due to defective genital papillae but possessed functional mature sperm. All defects in the scd-/- mutants could be rescued by ubiquitous transgenic overexpression of scd. In conclusion, our study demonstrates that scd is indispensable for maintaining lipid homeostasis and development of secondary sexual characteristics in zebrafish.

Revealing the process of storage protein rebalancing in high quality protein maize by proteomic and transcriptomic

Quality protein maize(QPM)(Zea mays L.) varieties contain enhanced levels of tryptophan and lysine, exhibiting improved nutritive value for humans and livestock. However, breeding QPM varieties remains challenging due to the complex process of rebalancing storage protein. This study conducted transcriptome and proteome analyses to investigate the process of storage proteins rebalancing in opaque2(o2) and QPM. We found a weak correlation between the transcriptome and proteome, suggesting a significant modulating effect of post-transcriptional events on non-zein protein abundances in Mo17o2 and QPM. These results highlight the advantages of proteomics. Compared with Mo17, 672 differentially expressed proteins(DEPs) were identified both in Mo17o2 and QPM, and several of them were associated with storage protein, starch, and amino acid synthesis. We identified 178 non-zeins as DEPs in Mo17o2 and QPM kernels. The up-regulated non-zein DEPs were enriched in lysine, tryptophan, and methionine, which affected the protein quality. Co-expression network analysis identified regulators of storage protein synthesis in QPM, including O2,PBF1, and several transcription factors. Our results revealed how storage protein rebalancing occurs and identified nonzein DEPs that may facilitate superior-quality QPM breeding.