Mitochondrial localization of ORF346 causes pollen abortion in alloplasmic male sterility

The Nsa cytoplasmic male sterility(CMS) system confers stable male sterility and offers great potential for production of hybrid seeds in oilseed rape. However, genes responsible for male sterility in Nsa CMS have not been identified. By mitochondrial genome sequencing of Nsa CMS and its maintainer line,we identified in an Nsa CMS line several chimeric genes encoding hypothetical proteins harboring transmembrane domains. One novel chimeric gene orf346 showed high identity with cox1 at the 50 terminal region and was co-transcribed with nad3 and rps12 genes. Transgenic plants of orf346 fused with or without mitochondrial targeting peptide conferred complete male sterility in Arabidopsis. ORF346 was mitochondrion-localized. Expression of orf346 in Escherichia coli inhibited bacterial growth, with excessive accumulation of reactive oxygen species and decreased ATP content. These results reveal a link between the newly identified mitochondrial gene orf346 and the abortion of Nsa CMS. Inadequate energy supply and excessive accumulation of reactive oxygen species may account for pollen abortion in Nsa CMS plants.

A lignified-layer bridge controlled by a single recessive gene is associated with high pod-shatter resistance in Brassica napus L.

Pod shattering causes severe yield loss in rapeseed(Brassica napus L.)under modern agricultural practice.Identification of highly shatter-resistant germplasm is desirable for the development of rapeseed cultivars for mechanical harvesting.In the present study,an elite line OR88 with strong shatter resistance and a lignified-layer bridge(LLB)structure was identified.The LLB structure was unique to OR88 and co-segregated with high pod-shatter resistance.The LLB structure is differentiated at stage 12 of gynoecium development without any gynoecium defects.Genetic analysis showed that LLB is controlled by a single recessive gene.By BSA-Seq and map-based cloning,the resistance gene location was delimited to a0.688 Mb region on chromosome C09.Transcriptome analysis suggested Bn TCP8.C09 as the gene responsible for LLB.The expression of Bn TCP.C09 was strongly downregulated in OR88,suppressing cell proliferation in the pod valve margin.KASP markers linked to the candidate gene were developed.This pod shatter-resistant line could be used in rapeseed breeding programs by direct transfer of the gene with the assistance of the DNA markers.

Complete organelle genomes of Sinapis arvensis and their evolutionary implications

Sinapis arvensis,belonging to the genus Sinapis of the family Brassicaceae,has good agronomic characters that make it a valuable genetic resource for crop improvement and is a cytoplasmic source of heterologous cytoplasmic male sterility(CMS).In addition,S.arvensis has played an important role in the evolution of the six major cultivated Brassica species involved in the triangle of U.Using next-generation sequencing,we assembled and revealed the gene composition of S.arvensis cytoplasmic genome.The chloroplast genome comprises 153,590 bp,with 112 individual genes,including 4 r RNA,29 t RNA,and 79 proteincoding genes.The mitochondrial genome comprises 240,024 bp,with 54 genes,including 18 t RNA,three r RNA and 33 protein-coding genes.Genome structure and evolutionary analysis indicated that the sequences of the S.arvensis organellar genomes were more similar to those of Brassica nigra and B.carinata than to those of other Brassicaceae species.Four mitochondrial open reading frames displaying chimeric structural features and encoding hypothetical proteins with transmembrane domains may account for the infertility of Nsa CMS previously derived from somatic cell hybridization between B.napus and S.arvensis.These results will not only contribute to utilize the germplasm resource of S.arvensis,and comprehend the evolution of organelle genomes within the Brassicaceae family,but also help to identify genes conditioning the alloplasmic male sterility of Nsa CMS in B.napus.

Identification of compact germplasm resources suitable for high density cultivation in Brassica napus L.

Branch angle is an important plant architecture trait and is considerably important for the ideal plant architecture of high density cultivation of rapeseed. In this study,12 representative rapeseed lines were first measured for whole-plant branch angles using image processing technology. Top branch angle (TBA) and basal branch angle (BBA) were significantly different in each line in which TBA was greater than BBA. Then, 156 rapeseed germplasm lines were evaluated for TBA and BBA in main cultivated regions of the Yangtze River Basin (Zunyi; Wuhan; Lu'an). In these three environments, the TBA and BBA of the rapeseed germplasm sources varied, in which ranges of 33.44°-73.17°and 18.73°-59.11°, 29.66°-61.91°and 22.32°-60.16°, and 28.47°-76.32°and 22.98°-72.68°, respectively. ANOVA showed that TBA and BBA were susceptible to environmental and had broadsense heritability (H2) values of 89.09%and 87.40%, respectively. Interactions of genotype by environment were also significant. There was diversity in branch angle among rapeseed genotypes. GGE biplot analysis showed that compact accession lines (Nilla, Purler and AV-SAPPHI) were the most desirable compact genotypes in all environments. Stability analysis showed that the genotype with the most minimum branch angle was stable for compact plant architecture.

Establishment of new convenient two-line system for hybrid production by targeting mutation of OPR3 in allopolyploid Brassica napus

The two-line pollination control system,which usually depends on the utilization of thermosensitive or photoperiod genic male-sterile lines,has been widely used in various crops.However,this system is susceptible to instability issues caused by uncontrollable weather fluctuations.A stable and handy two-line pollination control system is highly desirable in many crop species for heterosis exploitation.Oxophytodienoic acid reductase 3(OPR3)was proven to be involved in jasmonate biosynthesis.In the present study,CRISPR/Cas9(Clustered Regularly Interspaced Short Palindromic Repeat)was utilized to mutate two OPR3 homologs in Brassica napus.After two OPR3 homologs were simultaneously mutated,mutants exhibited complete male sterility,and fertility could be easily restored by exogenous MeJA treatment.Hybrids produced from crosses between the opr3 sterile lines and normal varieties exhibited heterosis.This new two-line system based on OPR3 mutation provides higher stability and convenience than traditional systems.By using exogenous MeJA treatment to restore fertility,the system enables more precise control of male fertility transition,which has great potential to significantly contribute to the maneuverable production of hybrid seeds in rapeseed as well as other Brassica species crops.

An efficient approach for obtaining plant organelle genomes

Plant organelle(plastid and mitochondrial)genomes contain substantial information for plant evolution and adaptation.Therefore,it’s important to reveal plant whole-genome sequences including plastid and mitochondrial genomes.To decode these sequences,it is required to efficiently separate organelle genomic DNA from nucleus genome,which is difficult and laborious.In this study,an efficient procedure was established to obtain plant organelle genomes without extraction of plastid and mitochondria.Organelle DNA was extracted from three materials including Sinapis arvensis var.‘Yeyou 18’,a cytoplasmic male sterile line(Nsa CMS)and its corresponding maintainer line‘Zhongshuang 4’.DNA was sequenced by Roche 454 FLXt and Illumina Miseq platforms.Organelle genomes were assembled using the generated reads and public organelle genome sequences.This research presented a procedure that efficiently assembled organelle genomes and subsequent fill gaps by extending the consensus contig terminals.This method enabled us to assemble plant plastid and mitochondrial genomes simultaneously.The obtained organelle genomes could accelerate understanding of mitochondrial rearrangements and laid a foundation for further study of Sinapis arvensis evolution and sterility gene of Nsa CMS.

QTL mapping for yield components of Brassica napus L. using double haploid and immortalized F_2 populations

Effective silique number per plant(ESN), seeds per silique(SS), thousand seeds weight(TSW), silique length(SL) and silique density(SD) are important seed yield potential determinant traits in rapeseed(Brassica napus L.), which are controlled by quantitative trait loci(QTL). Mapping QTL to delimited chromosomal region offers an effective method for genetic dissection of these traits. A set of 96 double haploid(DH) lines were developed by crossing 2 Brassica napus lines R1 and R2, and an immortalized F_2(IF_2) population containing 124 combinations was developed by crossing those DH lines. DH populations were planted at 2 locations for 2 years and IF_2 populations were planted in 2 locations for 1 year. Based on the established 2,217.2 cM length high density genetic map, 42 QTLs were identified, with 26 QTLs detected repeatedly in different environments or populations, including 8 for SL, 7 for TSW, 4 for ESN, 4 for SS and 3 for SD. Among these identified QTLs, 3, 4, 1, 1 and 3 QTLs were considered as major QTLs for SL, TSW, ESN, SS and SD, respectively. In addition, 2 QTLs on A9 chromosome which control multiple traits were identified. These results warrant further study of fine mapping for yield and yield components.

Genetic relationship of rapeseed yield-related traits revealed by elite lines under multiple environments

To provide a theoretical basis for further improvement of Brassica napus yield, additive dominance with additive - by - additive epistatic effects （ ADAA） genetic model and a 6 X 8 partial dial- lel cross design were used to analyze the genetic effects and correlations of five yield related traits of 14 excellent Brassica napus parental lines and their 46 and F2 populations. The results showed that silique density （SD） , siliques per plant （SPP） , seeds per silique （SPS） and thousand - seed weight （TSW） exhibited not only additive and dominant effects, but also significant epistatic effects. The dominant effects of all five yield - related traits were obviously greater than their additive effects and epistatic effects. Yield per plant （YPP） showed significant genetic correlation with SD, SPP and SPS, and the main component of the genetic correlation was the dominance correlation. SPP and SPS both showed a significant negative correlation with TSW. The SD of rapeseed was genetically correlated with all three components of yield to a certain extent, and there were different components of genetic effects positively correlated with the three yield components, indicating that SD is a potential trait to reconcile the conflict between TSW and SPP as well as SPS.

液体活检在肺癌管理中新技术的发展和临床应用的拓展

Lung cancer is an exceedingly malignant tumor reported as having the highest morbidity and mortality of any cancer worldwide,thus posing a great threat to global health.Despite the growing demand for precision medicine,current methods for early clinical detection,treatment and prognosis monitoring in lung cancer are hampered by certain bottlenecks.Studies have found that during the formation and development of a tumor,molecular substances carrying tumor-related genetic information can be released into body fluids.Liquid biopsy(LB),a method for detecting these tumor-related markers in body fluids,maybe a way to make progress in these bottlenecks.In recent years,LB technology has undergone rapid advancements.Therefore,this review will provide information on technical updates to LB and its potential clinical applications,evaluate its effectiveness for specific applications,discuss the existing limitations of LB,and present a look forward to possible future clinical applications.Specifically,this paper will introduce technical updates from the prospectives of engineering breakthroughs in the detection of membrane-based LB biomarkers and other improvements in sequencing technology.Additionally,it will summarize the latest applications of liquid biopsy for the early detection,diagnosis,treatment,and prognosis of lung cancer.We will present the interconnectedness of clinical and laboratory issues and the interplay of technology and application in LB today.

Magnesium lithospermate B enhances the potential of human-induced pluripotent stem cell-derived cardiomyocytes for myocardial repair

Background:We previously reported that activation of the cell cycle in human-induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs)enhances their remuscularization capacity after human cardiac muscle patch transplantation in infarcted mouse hearts.Herein,we sought to identify the effect of magnesium lithospermate B(MLB)on hiPSC-CMs during myocardial repair using a myocardial infarction(MI)mouse model.Methods:In C57BL/6 mice,MI was surgically induced by ligating the left anterior descending coronary artery.The mice were randomly divided into five groups(n=10 per group);a MI group(treated with phosphate-buffered saline only),a hiPSC-CMs group,a MLB group,a hiPSC-CMs+MLB group,and a Sham operation group.Cardiac function and MLB therapeutic efficacy were evaluated by echocardiography and histochemical staining 4 weeks after surgery.To identify the associated mechanism,nuclear factor(NF)-κB p65 and intercellular cell adhesion molecule-1(ICAM1)signals,cell adhesion ability,generation of reactive oxygen species,and rates of apoptosis were detected in human umbilical vein endothelial cells(HUVECs)and hiPSC-CMs.Results:After 4 weeks of transplantation,the number of cells that engrafted in the hiPSC-CMs+MLB group was about five times higher than those in the hiPSC-CMs group.Additionally,MLB treatment significantly reduced tohoku hospital pediatrics-1(THP-1)cell adhesion,ICAM1 expression,NF-κB nuclear translocation,reactive oxygen species production,NF-κB p65 phosphorylation,and cell apoptosis in HUVECs cultured under hypoxia.Similarly,treatment with MLB significantly inhibited the apoptosis of hiPSC-CMs via enhancing signal transducer and activator of transcription 3(STAT3)phosphorylation and B-cell lymphoma-2(BCL2)expression,promoting STAT3 nuclear translocation,and downregulating BCL2-Associated X,dual specificity phosphatase 2(DUSP2),and cleaved-caspase-3 expression under hypoxia.Furthermore,MLB significantly suppressed the production of malondialdehyde and lactate dehydrogenase and the reduction in glutathione content induced by hypoxia in both HUVECs and hiPSC-CMs in vitro.Conclusions:MLB significantly enhanced the potential of hiPSC-CMs in repairing injured myocardium by improving endothelial cell function via the NF-κB/ICAM1 pathway and inhibiting hiPSC-CMs apoptosis via the DUSP2/STAT3 pathway.

MoO_(x) nanoclusters on Mo-doped TiO_(2) nanosheets with enhanced singlet oxygen generation and sulfide conversion abilities for photocatalytic aerobic oxidative desulfurization

Photocatalytic aerobic oxidation desulfurization(PAODS)is a promising and sustainable alternative to conventional,energyintensive desulfurization techniques for petroleum products.However,its development is greatly plagued by the low capability in generating highly reactive oxygen species and sluggish kinetics of sulfide oxidation of reported photocatalysts.Here we report a class of MoO_(x)nanocluster decorated on ultrathin Mo-doped TiO_(2)nanosheet(MoO_(x)/MoTiO)catalyst for efficiently facilitating the photocatalytic aerobic oxidation of sulfides.We demonstrate that MoO_(x)/MoTiO can not only promote the generation of highly reactive singlet oxygen(^(1)O_(2))but also enhance the aerobic conversion of sulfides,which leads to a record dibenzothiophene oxidation activity of 3.90 mmol g^(-1)h^(-1).The multiple experimental characterizations and density functional theory calculations collectively reveal that the doped-Mo sites can interact with the photogenerated excitons,enabling directly energy transfer generation of^(1)O_(2)through a new exciton modulation mechanism,and the coordination unsaturated MoO_(x)clusters play the role of co-catalyst to enhance the separation of charge carriers,and effectively catalyze the reaction between sulfides and1O_(2)to form sulfones.

A Kelch Motif-Containing Serine/Threonine Protein Phosphatase Determines the Large Grain QTL Trait in Rice

A thorough understanding of the genetic basis of rice grain traits is critical for the improvement of rice （Oryza sativa L.） varieties. In this study, we generated an F2 population by crossing the large-grain japonica cultivar CW23 with Peiai 64 （PA64）, an elite indica small-grain cultivar. Using QTL analysis, 17 QTLs for five grain traits were detected on four different chromosomes. Eight of the QTLs were newly-identified in this study. In particular, qGL3-1, a newly-identified grain length QTL with the highest LOD value and largest phenotypic variation, was fine-mapped to the 17 kb region of chromosome 3. A serine/threonine protein phosphatase gene encoding a repeat domain containing two Kelch motifs was identified as the unique candidate gene corresponding to this QTL. A comparison of PA64 and CW23 sequences revealed a single nucleotide substitution （C→A） at position 1092 in exon 10, resulting in replacement of Asp （D） in PA64 with Glu （E） in CW23 for the 364th amino acid. This variation is located at the D position of the conserved sequence motif AVLDT of the Kelch repeat. Genetic analysis of a near-isogenic line （NIL） for qGL3-1 revealed that the allele qGL3-1 from CW23 has an additive or partly dominant effect, and is suitable for use in molecular marker-assisted selection.

A terbium activated multicolour photoluminescent phosphor for luminescent anticounterfeiting

The multicolor photoluminescence(PL) of a Tb3+singly activated NaYGeO4 phosphor is prepared by solid state method.For the first time,it is reported that the PL color of Tb3+in NaYGeO4 phosphor can be tuned from green to blue and,finally,to red by controlling the Tb3+content.The optimal tricolor phosphors are NaYGeO4:0.01 mol% Tb3+(red),NaYGeO4:0.5 mol% Tb3+(blue) and NaYGeO4:5 mol% Tb3+(green),respectively.The PL properties of the NaYGeO4:Tb3+ phosphor were studied in details.The results reveal that the PL color modulations of the NaYGeO4:Tb3+phosphor are essentially due to the cross-relaxation effect of Tb3+,and a possible mechanism on the cross-relaxation is proposed.According to these unique features,a multicolor PL image was fabricated based on the NaYGeO4:Tb3+ phosphor for potential applications in luminescent anticounterfeiting.

Ferromagnetism in sp^(2) carbon

The bulk,pristine sp^(2) carbons,such as graphite,carbon nanotubes,and graphene,are usually assumed to be typical diamagnetic materials.However,over the past two decades,there have been many reports about the ferromagnetism in these sp^(2) carbon materials,which have attracted intense interest for basic research and potential applications.In this review,we focus on the evidence and developments of the emergent ferromagnetism in sp^(2) carbon revealed by nine kinds of experimental methods:magnetic force microscopy(MFM),magnetization measurements with physical property measurement system(PPMS),X-ray magnetic circular dichroism(XMCD),scanning tunneling microscopy(STM),miniaturized magnetic particle inspection(MPI),anomalous Hall effect(AHE),mechanical deflection of carbon nanotube cantilevers,magnetoresistance,and spin-related devices(spin field effect transistor and spin memory).The advantages,conclusions,challenges,and future of these methods are discussed.The ferromagnetism in sp^(2) carbon will open a door to explore exotic physical phenomena and lay the basis for the development of integrated circuit of spintronics,which is fundamentally different from charge-based conventional electronics.

Representation Learning with Deconvolution for Multivariate Time Series Classification and Visualization

We propose a new model based on the convolutional networks and SAX(Symbolic Aggregate Approximation)discretization to learn the representation for multivariate time series.The deep neural networks has excellent expressiveness,which is fully exploited by the convolutional networks with means of unsupervised learning.We design a network structure to obtain the cross-channel correlation with means of convolution and deconvolution,the pooling operation is utilized to perform the dimension reduction along each position of the channels.Discretization which based on the Symbolic Aggregate Approximation is applied on the feature vectors to extract the bag of features.We collect two different representations from the convolutional networks,the compression from bottle neck and the last convolutional layers.We show how these representations and bag of features can be useful for classification.We provide a full comparison with the sequence distance based approach on the standard datasets to demonstrate the effectiveness of our method.We further build the Markov matrix according to the discretized representation abstracted from the deconvolution,time series is visualized to complex networks through Markov matrix visualization,which show more class-specific statistical properties and clear structures with respect to different labels.