CRISPR/Cas9-mediated mutagenesis of CiBG1 decreased seed size and promoted seed germination in watermelon

Abscisic acid(ABA)is a critical regulator of seed development and germination.β-glucosidases(BGs)have been suggested to be contributors to increased ABA content because they catalyze the hydrolysis of ABA-glucose ester to release free ABA.However,whether BGs are involved in seed development is unclear.In this study,a candidate gene,CiBG1,in watermelon was selected for targeted mutagenesis via the CRISPR/Cas9 system.Seed size and weight were significantly reduced in the Clbg1-mutant watermelon lines,which was mainly attributed to decreased cell number resulting from decreased ABA levels.A transcriptome analysis showed that the expression of 1015 and 1429 unique genes was changed 10 and 18 days after pollination(DAP),respectively.Cytoskeleton-and cell cycle-related genes were enriched in the differentially expressed genes of wild type and Clbg1-mutant lines during seed development.Moreover,the expression of genes in the major signaling pathways of seed size control was also changed.In addition,seed germination was promoted in the Cibg1-mutant lines due to decreased ABA content.These results indicate that ClBG1 may be critical for watermelon seed size regulation and germination mainly through the modulation of ABA content and thereby the transcriptional regulation of cytoskeleton-,cell cycle-and signaling-related genes.Our results lay a foundation for dissecting the molecular mechanisms of controlling watermelon seed size,a key agricultural trait of significant economic importance.

Correction: The NAC transcription factor ClNAC68 positively regulates sugar content and seed development in watermelon by repressing ClINV and ClGH3.6

In this article 1,author name Yanping Wang was incorrectly tagged as corresponding author.Prof.Wang should be the co-first author,instead of the corresponding author.The original article has been corrected.

Reconstructions of velocity distributions from fast-ion D-alpha(FIDA)measurements on EAST

2D fast-ion velocity-space distributions have been reconstructed from two-view fast-ion D-alpha(FIDA)measurements on experimental advanced superconducting tokamak(EAST).To make up for the sparse data and incomplete velocity-space coverage with the dual-view,we use nonnegativity and null-measurements as prior information to reconstruct the velocity distribution in experiments with co-and counter-current neutral beam injection.An improved reconstructed fast-ion distribution is achieved by combining the existing O-and B-port FIDA measurements with the proposed A-port FIDA view.To further improve the reliability of FIDA-based reconstructions on EAST,based on real multi-view FIDA measurements on EAST in the near future,various bases will be studied further.

The NAC transcription factor CINAC68 positively regulates sugar content and seed development in watermelon by repressing CIINV and CIGH3.6

NAC(NAM,ATAF1/2,and CUC2)transcription factors play important roles in fruit ripening and quality.The watermelon genome encodes 80 NAC genes,and 21 of these NAC genes are highly expressed in both the flesh and vascular tissues.Among these genes,CINAC68 expression was signi fi cantly higher in flesh than in rind.However,the intrinsic regulatory mechanism of CINAC68 in fruit ripening and quality is still unknown.In this study,we found that ClNAC68 is a transcriptional repressor and that the repression domain is located in the C-terminus.Knockout of CINAC68 by the CRISPR-Cas9 system decreased the soluble solid content and sucrose accumulation in mutant flesh.Development was delayed,germination was inhibited,and the IAA content was signi ficantly decreased in mutant seeds.Transcriptome analysis showed that the invertase gene CUNV was the only gene involved in sucrose metabolism that was upregulated in mutant flesh,and expression of the indole-3-acetic acid-amido synthetase gene CIGH3.6 in the IAA signaling pathway was also induced in mutant seeds.EMSA and dual-luciferase assays showed that CINAC68 directly bound to the promoters of CUNV and CIGH3.6 to repress their expression.These results indicated that CINAC68 positively regulated sugar and IAA accumulation by repressing CUNV and CIGH3.6.Our findings provide new insights into the regulatory mechanisms by which NAC transcription factors affect fruit quality and seed development.

Evaluation of NBI absorption and fast ion stored energy to improve thermal energy confinement time calculation in EAST

The absorption of neutral beam power and the fast ion stored energy in EAST plasmas with neutral beam injection(NBI)is analyzed to improve the calculation of thermal energy confinement time.The neutral beam power absorption and fast ion stored energy are systematically calculated using the TRANSP code,through the investigation of global parameters including plasma current,line averaged density and beam energy.Results have shown that scaling laws for the NBI absorption coefficient and fast ion energy rate are obtained through statistical analysis.A comparison of the confinement improvement factor H98y2 with these new scaling laws against those assuming fixed coefficients is given.

Differentiation between two healthcare concepts:Person-centered and patient-centered care

1.Background Engel's biopsychosocial model,which was constructed in 1977,was considered a momentous breakthrough from the traditional biomedical model of medicine that regards the person as a disease-carrier requiring diagnosis and treatment[1]towards recognition of the whole person and furnish customized care.This model argues that health should not be considered solely on biological factors but on a combination of biological,psychological,and social factors[2].This idea of considering individuals as a whole and respecting their rights to self-determination has led to a shift in the care model from disease-centered care to patient-centered or person-centered care.The latter two care models are both abbreviated as PCC.Ensuring that people are involved in and central to the healthcare process is now recognized as a key component in developing high-quality care.Studies have revealed that both concepts of care can help improve people's health condition and lower health service burdens.Thus,countries and healthcare organizations have been paying close attention to the patients'concerns.The National Health Service in England has made person-centered care as one of its seven core principles[3].

ClSnRK2.3 negatively regulates watermelon fruit ripening and sugar accumulation

Watermelon(Citrullus lanatus) as non-climacteric fruit is domesticated from the ancestors with inedible fruits. We previously revealed that the abscisic acid(ABA) signaling pathway gene ClSnRK2.3 might infuence watermelon fruit ripening. However,the molecular mechanisms are unclear. Here,we found that the selective variation of ClSnRK2.3 resulted in lower promoter activity and gene expression level in cultivated watermelons than ancestors, which indicated ClSnRK2.3 might be a negative regulator in fruit ripening. Overexpression (OE) of ClSnRK2.3 significantly delayed watermelon fruit ripening and suppressed the accumulation of sucrose, ABA and gibberellin GA4. Furthermore,we determined that the pyrophosphate-dependent phosphofructokinase(ClPFP1) in sugar metabolism pathway and GA biosynthesis enzyme GA20 oxidase(Cl GA20ox) could be phosphorylated by ClSnRK2.3 and thereby resulting in accelerated protein degradation in OE lines and finally led to low levels of sucrose and GA4. Besides that, ClSnRK2.3 phosphorylated homeodomain-leucine zipper protein(ClHAT1) and protected it from degradation to suppress the expression of the ABA biosynthesis gene 9’-cis-epoxycarotenoid dioxygenase 3(Cl NCED3). These results indicated that ClSnRK2.3 negatively regulated watermelon fruit ripening by manipulating the biosynthesis of sucrose, ABA and GA4. Altogether, these findings revealed a novel regulatory mechanism in non-climacteric fruit development and ripening.

Resonator fiber optic gyros with light time-division input and multiplexing output in clockwise and counterclockwise directions

A resonator fiber optic gyro with the light time-division input and multiplexing output(TDM-RFOG)in the clockwise(CW)and counterclockwise(CCW)directions is proposed.The light time-division input in the CW and CCW directions can effectively suppress the backscattering induced noise.The TDM-RFOG is implemented with the 2×2 Mach–Zehnder interferometer(MZI)optical switch.The response time of the fiber ring resonator is analyzed,and it is demonstrated by experiments that light time-division input in the CW and CCW directions can reduce the backscattering induced noise.The suppression effectiveness of backscattering induced noise in the TDM-RFOG is determined by the extinction ratio of the optical switch,so a closed loop is designed to adjust the phase shift difference between the two arms of the MZI optical switch to control the extinction ratio.The method using two arms of the MZI optical switch with twin 90°polarization-axis rotated splices is proposed to make the extinction ratio along both slow and fast axes greater than-70 d B.

Natural variation in the NAC transcription factor NONRIPENING contributes to melon fruit ripening

ThThe NAC transcription factor NONRIPENING(NOR)is a master regulator of climacteric fruit ripening.Melon(Cucumis melo L.)has climacteric and nonclimacteric fruit ripening varieties and is an ideal model to study fruit ripening.Two natural CmNAC-NOR variants,the climacteric haplotype CmNAC-NOR^(S,N) and the non-climacteric haplotype CmNAC-NOR^(A,S),have effects on fruit ripening;however,their regulatory mechanisms have not been elucidated.Here,we report that a natural mutation in the transcriptional activation domain of CmNAC-NORS,Ncontributes to climacteric melon fruit ripening.CmNAC-NOR knockout in the climacteric-type melon cultivar“BYJH”completely inhibited fruit ripening,while ripening was delayed by 5-8 d in heterozygous cmnac-nor mutant fruits.CmN AC-NOR directly activated carotenoid,ethylene,and abscisic acid biosynthetic genes to promote fruit coloration and ripening.Furthermore,CmNAC-NOR mediated the transcription of the“CmNAC-NOR-CmNAC73-CmCWINV2”module to enhance flesh sweetness.The transcriptional activation activity of the climacteric haplotype CmNAC-NORS,Non these target genes was significantly higher than that of the nonclimacteric haplotype CmNAC-NOR^(A,S).Moreover,CmNAC-NORS,Ncomplementation fully rescued the non-ripening phenotype of the tomato(Solanum lycopersicum)cr-nor mutant,while CmNAC-NOR^(A,S) did not.Our results provide insight into the molecular mechanism of climacteric and non-climacteric fruit ripening in melon.

A Novel Single-Nucleotide Mutation in a CLAVATA3 Gene Homolog Controls a Multilocular Silique Trait in Brassica rapa L.

Dear Editor, Brassica rapa is an important crop and a model plant for studying Brassica genome evolution and gene function. The silique of B. rapa derived from the gynoecium nor- mally consists of two carpels that are separated by a false septum, and thus has two Iocules （bilocular）. Multilocular （more than two carpels） lines of B. rapa have been discov- ered in nature and are considered advantageous because the multilocular type could potentially increase the seed yield by producing more seeds per silique than the biloc- ular type. However, the molecular basis of the important seed vield-related trait is ooorlv understood.