The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the ...The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the SCF complex and degraded by the 26S protease accounts for the bulk of the available self-incompatibility studies.In this study,15 ClSKP1s from the‘Xiangshui'lemon genome and ubiquitome exist in the same SKP1 conserved domain(CD)as SKP1s in other species.The q PCR results showed that SKP1-6 and SKP1-14 have tissue expression patterns specific for expression in pollen.In addition,SKP1-6 and SKP1-14 in the stigma,style and ovary were significantly upregulated after self-pollination compared to those after cross-pollination.A subcellular location showed that SKP1-6 and SKP1-14 were located in the nucleus.In addition,yeast two-hybrid(Y2H)assays,bimolecular fluorescence complementation(BiFC)and luciferase complementation imaging(LCI)assays showed that SKP1-6 interacted with F-box1,F-box33,F-box34,F-box17,F-box19,Cullin1-2 and 26S proteasome subunit 4 homolog A(26S PS4HA).SKP1-14 interacted with F-box17,F-box19,F-box35,Cullin1-2 and 26S PS4HA.The interaction of Cullin1-2 and the F-box with SKP1 as a bridge was verified by a yeast three-hybrid experiment.The ability of S3-RNase to inhibit pollen and pollen tube growth and development was assessed using in vitro pollen co-culture experiments with recombinant S3-RNase proteins.Overall,this study provides important experimental evidence and theoretical basis for understanding the mechanism of self-incompatibility in plants by revealing the key role of the SCF complex in‘Xiangshui'lemon,which is bridged by ClSKP1-6,in self-incompatibility.The results of this study are of great significance for the future indepth exploration of the molecular mechanism of the SCF complex and its wide application in the self-incompatibility of plants.展开更多
The width of rice leaves determines the size of the photosynthetic area.Optimizing rice leaf width can improve the photosynthetic rate,thereby increasing rice yield.In this study,a genome-wide association study(GWAS)w...The width of rice leaves determines the size of the photosynthetic area.Optimizing rice leaf width can improve the photosynthetic rate,thereby increasing rice yield.In this study,a genome-wide association study(GWAS)was conducted by 225 rice germplasm resources to explore the genetic basis of rice flag leaf width(FLW).We identified nine QTLs associated with FLW(qFLWs),with phenotypic contribution rates ranging from 3.17%to 14.37%.Near-isogenic lines(NILs)were developed for fine-mapping of qFLW11,and the function of FLW11 was further verified.We narrowed down q FLW11 to an 87-kb interval,which contains five genes.展开更多
[Objectives]This study was conducted to comprehensively understand the changes in gene expression of plants under environmental stress during different growth and development stages.[Methods]The effects of continuous ...[Objectives]This study was conducted to comprehensively understand the changes in gene expression of plants under environmental stress during different growth and development stages.[Methods]The effects of continuous cropping on the roots and leaves of Polygonatum sibiricum were investigated using transcriptome sequencing.Normally-grown first crop P.sibiricum was used as the control group,while continuous cropping plants served as the treatment group.Transcriptomic differences in roots and leaves under different conditions were compared.[Results]The leaf materials of first crop and continuous cropping P.sibiricum(CCLZ vs FCLZ)showed 21916 differentially expressed genes(DEGs),while the root materials of first crop and continuous cropping P.sibiricum(CCRZ vs FCRZ)exhibited 12726 DEGs(the lowest DEG count)(12726).Among them,1896 DEGs were common.GO enrichment analysis revealed that DEGs were mainly enriched in metabolism,cell wall degradation,and pathogen defense.KEGG enrichment analysis indicated that DEGs in CCLZ vs FCLZ and CCRZ vs FCRZ primarily affected hormone signal transduction and pathogen interaction pathways.[Conclusions]This study preliminarily elucidate the regulatory mechanisms in the roots and leaves of continuous cropping P.sibiricum at the molecular level,providing reference for research on its adaptation to continuous cropping.展开更多
Temperature plays a crucial role in shaping microbial ecosystems during anaerobic digestion.However,the specific microbial communities and their functions across a wide temperature range still remain elusive.This stud...Temperature plays a crucial role in shaping microbial ecosystems during anaerobic digestion.However,the specific microbial communities and their functions across a wide temperature range still remain elusive.This study employed a genome-centric metagenomic approach to explore microbial metabolic pathways and synergistic networks at temperatures of 35,44,53,55,and 65℃.A total of 84 metagenome assembled genomes(MAGs)were assembled,with over 65%of these MAGs corresponding to novel bacterial and archaeal species(including Firmicutes,Chloroflexota,Bacteroidia and Methanobacteriota),greatly enhancing our current comprehension anaerobic digestion process.Notably,functional annotation identified that 44_bin.2(Methanothrix_A sp.001602645)harbors enzymes associated with hydrogenotrophic metabolism.Additionally,this microorganism exhibited diverse metabolic capabilities at 44℃,a temperature commonly employed in industrial practice yet less explored in bench studies.Consequently,it implies a promising potential for conducting anaerobic digestion at a moderate thermophilic temperature,as opposed to the conventional mesophilic range.The microorganism exhibited a variety of metabolic capabilities at 44℃,a temperature frequently employed in industrial applications but underexplored in laboratory investigations.The findings suggest that anaerobic digestion carried out at moderate thermophilic temperatures may have a higher potential for methane production.展开更多
In this paper,a standardized analysis method is established for identifying meat quality-related genes in Ordos finewool sheep using transcriptome sequencing data.A meticulously standardized approach is utilized to in...In this paper,a standardized analysis method is established for identifying meat quality-related genes in Ordos finewool sheep using transcriptome sequencing data.A meticulously standardized approach is utilized to investigate the genetic determinants of meat quality in Ordos fine-wool sheep through transcriptome sequencing analysis.Muscle samples from the longissimus dorsi of one-year-old sheep are collected under controlled conditions,and key texture properties—hardness,elasticity,and chewiness—are measured to categorize samples into high-and low-textural-value groups.Genes significantly associated with meat quality traits are identified through standardized RNA extraction,high-throughput sequencing,and differential gene expression analysis.Functional enrichment analysis reveals their involvement in biological processes such as extracellular matrix organization and metabolic pathways.The findings underscore the pivotal role of standardization in meat quality research,laying a solid scientific foundation for future research on meat quality improvement and molecular breeding.展开更多
AIM： TO investigate the protein profile of human hepatocarcinoma cell line SMMC-7721, to analyze the specific functions of abundant expressed proteins in the processes of hepatocarcinoma genesis, growth and metastasi...AIM： TO investigate the protein profile of human hepatocarcinoma cell line SMMC-7721, to analyze the specific functions of abundant expressed proteins in the processes of hepatocarcinoma genesis, growth and metastasis, to identify the hepatocarcinoma-specific biomarkers for the early prediction in diagnosis, and to explore the new drug targets for liver cancer therapy. METHODS： Total proteins from human hepatocarcinoma cell line SMMC-7721 were separated by two-dimensional electrophoresis （2DE）. The silver-stained gel was analyzed by 2DE software Image Master 2D Elite. Interesting protein spots were identified by peptide mass fingerprinting based on matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and database searching. RESULTS： We obtained protein profile of human hepatocarcinoma cell line SMMC-7721. Among the twenty-one successfully identified proteins, mitofilin, endoplasmic reticulum protein ERp29, ubiquinol-cytochrome C reductase complex core protein I, peroxisomal enoyl CoA hydratase, peroxiredoxin-4 and probable 3-oxoacid CoA transferase 1 precursor were the six novel proteins identified in human hepatocarcinoma cells or tissues. Specific functions of the identified heat-shock proteins were analyzed in detail, and the results suggested that these proteins might promote tumorigenesis via inhibiting cell death induced by several cancer-related stresses or via inhibiting apoptosis at multiple points in the apoptotic signal pathway. Other identified chaperones and cancer-related proteins were also analyzed.CONCLUSION： Based on the protein profile of SMMC-7721 cells, functional analysis suggests that the identified chaperones and cancer-related proteins have their own pathways to contribute to the tumorigenesis, tumor growth and metastasis of liver cancer. Furthermore, proteomic analysis is indicated to be feasible in the cancer study.展开更多
This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpot...This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame （ORF） encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point （pI） of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD （flavinadenine dinucleotide）/NAD（P） （nicotinamide adenine dinucleotide phosphate）-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco （cv. Wisconsin 38） expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.展开更多
This study presents an analysis of the spectral characteristics of remote sensing reflectance(Rrs)in northwestern South China Sea based on the in situ optical and water quality data for August 2018.Rrswas initially di...This study presents an analysis of the spectral characteristics of remote sensing reflectance(Rrs)in northwestern South China Sea based on the in situ optical and water quality data for August 2018.Rrswas initially divided into four classes,classes A to D,using the max-classification algorithm,and the spectral properties of whole Rrs were characterized using the empirical orthogonal function(EOF)analysis.Subsequently,the dominant factors in each EOF mode were determined.The results indicated that more than 95%of the variances of Rrs are partly driven by the back-scattering characteristics of the suspended matter.The initial two EOF modes were well correlated with the total suspended matter and back-scattering coefficient.Furthermore,the first EOF modes of the four classes of Rrs(A-D Rrs-EOF1)significantly contributed to the total variances of each Rrs class.In addition,the correlation coefficients between the amplitude factors of class A-D Rrs-EOF1 and the variances of the relevant water quality and optical parameters were better than those of the unclassified ones.The spectral shape of class ARrs-EOF1 was governed by the absorption characteristic of chlorophyll a and colored dissolved organic matter(CDOM).The spectral shape of class B Rrs-EOF1 was governed by the absorption characteristic of CDOM since it exhibited a high correlation with the absorption coefficient of CDOM(ag(λ)),whereas the spectral shape of class C Rrs-EOF1 was governed by the back-scattering characteristics but not affected by the suspended matter.The spectral shape of class D Rrs-EOF1 exhibited a relatively good correlation with all the water quality parameters,which played a significant role in deciding its spectral shape.展开更多
To our knowledge,few reports on Demodex studied at the molecular level are available at present.In this study our group,for the first time,cloned,sequenced and analyzed the chitin synthase(CHS) gene fragments of Demod...To our knowledge,few reports on Demodex studied at the molecular level are available at present.In this study our group,for the first time,cloned,sequenced and analyzed the chitin synthase(CHS) gene fragments of Demodex folliculorum,Demodex brevis,and Demodex canis(three isolates from each species) from Xi'an China,by designing specific primers based on the only partial sequence of the CHS gene of D.canis from Japan,retrieved from GenBank.Results show that amplification was successful only in three D.canis isolates and one D.brevis isolate out of the nine Demodex isolates.The obtained fragments were sequenced to be 339 bp for D.canis and 338 bp for D.brevis.The CHS gene sequence similarities between the three Xi'an D.canis isolates and one Japanese D.canis isolate ranged from 99.7% to 100.0%,and those between four D.canis isolates and one D.brevis isolate were 99.1%-99.4%.Phylogenetic trees based on maximum parsimony(MP) and maximum likelihood(ML) methods shared the same clusters,according with the traditional classification.Two open reading frames(ORFs) were identified in each CHS gene sequenced,and their corresponding amino acid sequences were located at the catalytic domain.The relatively conserved sequences could be deduced to be a CHS class A gene,which is associated with chitin synthesis in the integument of Demodex mites.展开更多
Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both...Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.展开更多
Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes ...Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.The gene expression profiles of CD11 b^(+)/Ly6 C^(intermediate)macrophages of the sham surgery mice,and the mice 4 h,24 h and 9 days after renal IRI were downloaded from the Gene Expression Omnibus database.Analysis of m RNA expression profiles was conducted to identify differentially expressed genes(DEGs),biological processes and pathways by the series test of cluster.Protein-protein interaction network was constructed and analysed to discover the key genes.A total of 6738 DEGs were identified and assigned to 20 model profiles.DEGs in profile 13 were one of the predominant expression profiles,which are involved in immune cell chemotaxis and proliferation.Signet analysis showed that Atp5 a1,Atp5 o,Cox4 i,Cdc42,Rac2 and Nhp2 were the key genes involved in oxidation-reduction,apoptosis,migration,M1-M2 differentiation,and proliferation of macrophages.RPS18 may be an appreciate reference gene as it was stable in macrophages.The identified DEGs and their enriched pathways investigate factors that may participate in the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.Moreover,the vital gene Nhp2 may involve the polarization of macrophages,which may be a new target to affect the process of AKI.展开更多
Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based ...Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based on bioinformatic analysis, we found that Arabidopsis genome actually encoded 17 OVATE domain-containing proteins. One of them, AtOFP19, has not been previously identified. Based on their amino acid sequence similarity, AtOFPs proteins can be divided into two groups. Most of the AtOFPs were located in nuclear, four of them were presented in chloroplast and the remaining two members appeared in cytoplasmic. A genome- wide microarray based gene expression analysis involving 47 stages of vegetative and reproductive development revealed that AtOFPs have diverse expression pattems. Investigation of proteins interaction showed that nine AtOFPs only interacted with TALE homeodomain proteins, which are fundamental regulators of plant meristem function and leaf development. Our work could provide important leads toward functional genomics studies of ovate family proteins, which may be involved in a previously unrecognized control mechanism in plant development展开更多
Traditional Global Sensitivity Analysis（GSA） focuses on ranking inputs according to their contributions to the output uncertainty.However,information about how the specific regions inside an input affect the output ...Traditional Global Sensitivity Analysis（GSA） focuses on ranking inputs according to their contributions to the output uncertainty.However,information about how the specific regions inside an input affect the output is beyond the traditional GSA techniques.To fully address this issue,in this work,two regional moment-independent importance measures,Regional Importance Measure based on Probability Density Function（RIMPDF） and Regional Importance Measure based on Cumulative Distribution Function（RIMCDF）,are introduced to find out the contributions of specific regions of an input to the whole output distribution.The two regional importance measures prove to be reasonable supplements of the traditional GSA techniques.The ideas of RIMPDF and RIMCDF are applied in two engineering examples to demonstrate that the regional moment-independent importance analysis can add more information concerning the contributions of model inputs.展开更多
Estimating the Probability Density Function（PDF） of the performance function is a direct way for structural reliability analysis,and the failure probability can be easily obtained by integration in the failure domai...Estimating the Probability Density Function（PDF） of the performance function is a direct way for structural reliability analysis,and the failure probability can be easily obtained by integration in the failure domain.However,efficiently estimating the PDF is still an urgent problem to be solved.The existing fractional moment based maximum entropy has provided a very advanced method for the PDF estimation,whereas the main shortcoming is that it limits the application of the reliability analysis method only to structures with independent inputs.While in fact,structures with correlated inputs always exist in engineering,thus this paper improves the maximum entropy method,and applies the Unscented Transformation（UT） technique to compute the fractional moments of the performance function for structures with correlations,which is a very efficient moment estimation method for models with any inputs.The proposed method can precisely estimate the probability distributions of performance functions for structures with correlations.Besides,the number of function evaluations of the proposed method in reliability analysis,which is determined by UT,is really small.Several examples are employed to illustrate the accuracy and advantages of the proposed method.展开更多
Reliability allocation of computerized numerical controlled(CNC)lathes is very important in industry.Traditional allocation methods only focus on high-failure rate components rather than moderate failure rate componen...Reliability allocation of computerized numerical controlled(CNC)lathes is very important in industry.Traditional allocation methods only focus on high-failure rate components rather than moderate failure rate components,which is not applicable in some conditions.Aiming at solving the problem of CNC lathes reliability allocating,a comprehensive reliability allocation method based on cubic transformed functions of failure modes and effects analysis(FMEA)is presented.Firstly,conventional reliability allocation methods are introduced.Then the limitations of direct combination of comprehensive allocation method with the exponential transformed FMEA method are investigated.Subsequently,a cubic transformed function is established in order to overcome these limitations.Properties of the new transformed functions are discussed by considering the failure severity and the failure occurrence.Designers can choose appropriate transform amplitudes according to their requirements.Finally,a CNC lathe and a spindle system are used as an example to verify the new allocation method.Seven criteria are considered to compare the results of the new method with traditional methods.The allocation results indicate that the new method is more flexible than traditional methods.By employing the new cubic transformed function,the method covers a wider range of problems in CNC reliability allocation without losing the advantages of traditional methods.展开更多
Statistical analysis was done on simultaneous wave and wind using data recorded by discus-shape wave buoy. The area is located in the southern Caspian Sea near the Anzali Port. Recorded wave data were obtained through...Statistical analysis was done on simultaneous wave and wind using data recorded by discus-shape wave buoy. The area is located in the southern Caspian Sea near the Anzali Port. Recorded wave data were obtained through directional spectrum wave analysis. Recorded wind direction and wind speed were obtained through the related time series as well. For 12-month measurements（May 25 2007-2008）, statistical calculations were done to specify the value of nonlinear auto-correlation of wave and wind using the probability distribution function of wave characteristics and statistical analysis in various time periods. The paper also presents and analyzes the amount of wave energy for the area mentioned on the basis of available database. Analyses showed a suitable comparison between the amounts of wave energy in different seasons. As a result, the best period for the largest amount of wave energy was known. Results showed that in the research period, the mean wave and wind auto correlation were about three hours. Among the probability distribution functions, i.e Weibull, Normal, Lognormal and Rayleigh, ＂Weibull＂ had the best consistency with experimental distribution function shown in different diagrams for each season. Results also showed that the mean wave energy in the research period was about 49.88 k W/m and the maximum density of wave energy was found in February and March, 2010.展开更多
This study analyzed the function of different glutathione S-transferase(GST)isoforms and detoxification metabolism responses in Manila clam,Ruditapes philippinarum,exposed to 4 kinds of polycyclic aromatic hydrocarbon...This study analyzed the function of different glutathione S-transferase(GST)isoforms and detoxification metabolism responses in Manila clam,Ruditapes philippinarum,exposed to 4 kinds of polycyclic aromatic hydrocarbons(PAHs)single,and their mixtures for 15 days under laboratory conditions.13 kinds of GSTs in R.philippinarum were classified,and the results of tissue distribution indicated that 12 kinds of GSTs(except GST sigma 3)expressed most in digestive glands.We detected the m RNA expression levels of aryl hydrocarbon receptor signaling pathway,and detoxification system in digestive glands of clams exposed to benzo[a]pyrene(Ba P),chrysene(CHR),benzo[a]anthracene(Ba A),benzo[b]fluoranthene(Bb F),and Ba P+CHR+Ba A+Bb F,respectively.Among these genes,we selected GST-sigma,GST-omega and GST-pi as potential indicators to Ba P;GST-sigma,GST-A and GST-rho to CHR;GST-pi,GST-sigma,GST-A,GST-rho and GST-microsomal to Ba A;GST-theta and GST-mu to Bb F;while GST-pi and GST-mu to the mixture of Ba P,CHR,Ba A and Bb F.Additionally,the bioaccumulation of PAHs in tissues increased remarkably over time,and showed an obvious doseeffect.Under the same concentration,the bioaccumulation in single exposure group was higher than that in mixture group,and the bioaccumulation of PAHs in tissues with different concentrations of stress was irregular.The results revealed the metabolic differences and bioaccumulation rules in clams exposed to four kinds of PAHs,and provided more valuable information for the PAHs risk assessment.展开更多
In order to make rail transit smart stations serve passengers better,the potential of smart stations should be optimized to reduce time cost,and relieve traffic congestion.In this paper,the construction of smart stati...In order to make rail transit smart stations serve passengers better,the potential of smart stations should be optimized to reduce time cost,and relieve traffic congestion.In this paper,the construction of smart stations based on the management experience of a subway station and the benefits of smart stations were analyzed and discussed.The construction of smart stations,as well as the key technologies for the construction of smart stations,will aid in the automation and intelligent management of subway stations.展开更多
An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorop...An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.展开更多
基金supported by grants from the National Natural Science Foundation of China(Grant No.31960585)Science and Technology Major Project of Guangxi(Grant No.Guike AA22068092)+1 种基金Guangxi Science and Technology Vanguard Special Action Project(Grant No.202204)State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources(Grant Nos.SKLCUSA-a201906,SKLCU-SA-c201901)。
文摘The SKP1 gene is an important component of the SCF(SKP1-Cullin1-F-box)complex and serves as a bridge connecting the F-box and Cullin1genes(F-box-SKP1-Cullin1).The pattern of S-RNase being ubiquitously labelled by the SCF complex and degraded by the 26S protease accounts for the bulk of the available self-incompatibility studies.In this study,15 ClSKP1s from the‘Xiangshui'lemon genome and ubiquitome exist in the same SKP1 conserved domain(CD)as SKP1s in other species.The q PCR results showed that SKP1-6 and SKP1-14 have tissue expression patterns specific for expression in pollen.In addition,SKP1-6 and SKP1-14 in the stigma,style and ovary were significantly upregulated after self-pollination compared to those after cross-pollination.A subcellular location showed that SKP1-6 and SKP1-14 were located in the nucleus.In addition,yeast two-hybrid(Y2H)assays,bimolecular fluorescence complementation(BiFC)and luciferase complementation imaging(LCI)assays showed that SKP1-6 interacted with F-box1,F-box33,F-box34,F-box17,F-box19,Cullin1-2 and 26S proteasome subunit 4 homolog A(26S PS4HA).SKP1-14 interacted with F-box17,F-box19,F-box35,Cullin1-2 and 26S PS4HA.The interaction of Cullin1-2 and the F-box with SKP1 as a bridge was verified by a yeast three-hybrid experiment.The ability of S3-RNase to inhibit pollen and pollen tube growth and development was assessed using in vitro pollen co-culture experiments with recombinant S3-RNase proteins.Overall,this study provides important experimental evidence and theoretical basis for understanding the mechanism of self-incompatibility in plants by revealing the key role of the SCF complex in‘Xiangshui'lemon,which is bridged by ClSKP1-6,in self-incompatibility.The results of this study are of great significance for the future indepth exploration of the molecular mechanism of the SCF complex and its wide application in the self-incompatibility of plants.
基金supported by the Zhejiang Provincial Natural Science Foundation,China(Grant No.LD24C130001)the National Natural Science Foundation of China(Grant Nos.W2412006 and 32372125)+3 种基金the Hainan Provincial Natural Science Foundation,China(Grant Nos.GHYF2025029 and YBXM2422)the Innovation Platform for Academicians of Hainan Province,China(Grant No.YSPTZX202502)the National Modern Agricultural Industry Technology System Project,China(Grant No.CARS-01-18)the Special Support Program of Chinese Academy of Agricultural Sciences(Grant Nos.NKYCLJ-C-2021-015 and CAAS-ZDRW202401)。
文摘The width of rice leaves determines the size of the photosynthetic area.Optimizing rice leaf width can improve the photosynthetic rate,thereby increasing rice yield.In this study,a genome-wide association study(GWAS)was conducted by 225 rice germplasm resources to explore the genetic basis of rice flag leaf width(FLW).We identified nine QTLs associated with FLW(qFLWs),with phenotypic contribution rates ranging from 3.17%to 14.37%.Near-isogenic lines(NILs)were developed for fine-mapping of qFLW11,and the function of FLW11 was further verified.We narrowed down q FLW11 to an 87-kb interval,which contains five genes.
基金Supported by Agricultural Science and Technology Innovation Fund of Hunan Province(XCNZ[2021]No.15)Loudi Science and Technology Innovation Program(LKF[2022]29)+1 种基金Applied Characteristic Discipline Construction Project of Hunan Province:Plant ProtectionPostgraduate Research and Innovation Project of Hunan University of Humanities,Science and Technology(ZSCX2022Y12).
文摘[Objectives]This study was conducted to comprehensively understand the changes in gene expression of plants under environmental stress during different growth and development stages.[Methods]The effects of continuous cropping on the roots and leaves of Polygonatum sibiricum were investigated using transcriptome sequencing.Normally-grown first crop P.sibiricum was used as the control group,while continuous cropping plants served as the treatment group.Transcriptomic differences in roots and leaves under different conditions were compared.[Results]The leaf materials of first crop and continuous cropping P.sibiricum(CCLZ vs FCLZ)showed 21916 differentially expressed genes(DEGs),while the root materials of first crop and continuous cropping P.sibiricum(CCRZ vs FCRZ)exhibited 12726 DEGs(the lowest DEG count)(12726).Among them,1896 DEGs were common.GO enrichment analysis revealed that DEGs were mainly enriched in metabolism,cell wall degradation,and pathogen defense.KEGG enrichment analysis indicated that DEGs in CCLZ vs FCLZ and CCRZ vs FCRZ primarily affected hormone signal transduction and pathogen interaction pathways.[Conclusions]This study preliminarily elucidate the regulatory mechanisms in the roots and leaves of continuous cropping P.sibiricum at the molecular level,providing reference for research on its adaptation to continuous cropping.
基金supported by the National Natural Science Foundation of China(No.52270138)。
文摘Temperature plays a crucial role in shaping microbial ecosystems during anaerobic digestion.However,the specific microbial communities and their functions across a wide temperature range still remain elusive.This study employed a genome-centric metagenomic approach to explore microbial metabolic pathways and synergistic networks at temperatures of 35,44,53,55,and 65℃.A total of 84 metagenome assembled genomes(MAGs)were assembled,with over 65%of these MAGs corresponding to novel bacterial and archaeal species(including Firmicutes,Chloroflexota,Bacteroidia and Methanobacteriota),greatly enhancing our current comprehension anaerobic digestion process.Notably,functional annotation identified that 44_bin.2(Methanothrix_A sp.001602645)harbors enzymes associated with hydrogenotrophic metabolism.Additionally,this microorganism exhibited diverse metabolic capabilities at 44℃,a temperature commonly employed in industrial practice yet less explored in bench studies.Consequently,it implies a promising potential for conducting anaerobic digestion at a moderate thermophilic temperature,as opposed to the conventional mesophilic range.The microorganism exhibited a variety of metabolic capabilities at 44℃,a temperature frequently employed in industrial applications but underexplored in laboratory investigations.The findings suggest that anaerobic digestion carried out at moderate thermophilic temperatures may have a higher potential for methane production.
基金funded by the 2023 Inner Mongolia Public Institution High-Level Talent Introduction Scientific Research Support Project,and the Ordos Municipal Science and Technology Major Special Project(Grant No.2022EEDSKJZDZX021).
文摘In this paper,a standardized analysis method is established for identifying meat quality-related genes in Ordos finewool sheep using transcriptome sequencing data.A meticulously standardized approach is utilized to investigate the genetic determinants of meat quality in Ordos fine-wool sheep through transcriptome sequencing analysis.Muscle samples from the longissimus dorsi of one-year-old sheep are collected under controlled conditions,and key texture properties—hardness,elasticity,and chewiness—are measured to categorize samples into high-and low-textural-value groups.Genes significantly associated with meat quality traits are identified through standardized RNA extraction,high-throughput sequencing,and differential gene expression analysis.Functional enrichment analysis reveals their involvement in biological processes such as extracellular matrix organization and metabolic pathways.The findings underscore the pivotal role of standardization in meat quality research,laying a solid scientific foundation for future research on meat quality improvement and molecular breeding.
基金Supported by the National Natural Science Foundation of China, No. 30370403the Key Project of Chinese Ministry of Education, No. 705046the Doctoral Foundation of Xi’an Jiaotong University, grants No. DFXJTU2005-05
文摘AIM： TO investigate the protein profile of human hepatocarcinoma cell line SMMC-7721, to analyze the specific functions of abundant expressed proteins in the processes of hepatocarcinoma genesis, growth and metastasis, to identify the hepatocarcinoma-specific biomarkers for the early prediction in diagnosis, and to explore the new drug targets for liver cancer therapy. METHODS： Total proteins from human hepatocarcinoma cell line SMMC-7721 were separated by two-dimensional electrophoresis （2DE）. The silver-stained gel was analyzed by 2DE software Image Master 2D Elite. Interesting protein spots were identified by peptide mass fingerprinting based on matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and database searching. RESULTS： We obtained protein profile of human hepatocarcinoma cell line SMMC-7721. Among the twenty-one successfully identified proteins, mitofilin, endoplasmic reticulum protein ERp29, ubiquinol-cytochrome C reductase complex core protein I, peroxisomal enoyl CoA hydratase, peroxiredoxin-4 and probable 3-oxoacid CoA transferase 1 precursor were the six novel proteins identified in human hepatocarcinoma cells or tissues. Specific functions of the identified heat-shock proteins were analyzed in detail, and the results suggested that these proteins might promote tumorigenesis via inhibiting cell death induced by several cancer-related stresses or via inhibiting apoptosis at multiple points in the apoptotic signal pathway. Other identified chaperones and cancer-related proteins were also analyzed.CONCLUSION： Based on the protein profile of SMMC-7721 cells, functional analysis suggests that the identified chaperones and cancer-related proteins have their own pathways to contribute to the tumorigenesis, tumor growth and metastasis of liver cancer. Furthermore, proteomic analysis is indicated to be feasible in the cancer study.
基金supported by the China Agriculture Research System (Sweetpotato)the National High-Tech Research and Development Project of China(2011AA100607 and 2012AA101204)
文摘This paper reported firstly successful cloning of lycopene ε-cyclase （lbLCYe） gene from sweetpotato, lpomoea batatas （L.） Lam. Using rapid amplification of cDNA ends （RACE）, lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame （ORF） encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point （pI） of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD （flavinadenine dinucleotide）/NAD（P） （nicotinamide adenine dinucleotide phosphate）-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco （cv. Wisconsin 38） expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.
基金The Key Projects of the Guangdong Education Department under contract No.2019KZDXM019the Fund of Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang)under contract No.ZJW-2019-08+2 种基金High-Level Marine Discipline Team Project of Guangdong Ocean University under contract No.002026002009the Guangdong Graduate Academic Forum Project under contract No.230420003the"First Class"discipline construction platform project in 2019 of Guangdong Ocean University under contract No.231419026。
文摘This study presents an analysis of the spectral characteristics of remote sensing reflectance(Rrs)in northwestern South China Sea based on the in situ optical and water quality data for August 2018.Rrswas initially divided into four classes,classes A to D,using the max-classification algorithm,and the spectral properties of whole Rrs were characterized using the empirical orthogonal function(EOF)analysis.Subsequently,the dominant factors in each EOF mode were determined.The results indicated that more than 95%of the variances of Rrs are partly driven by the back-scattering characteristics of the suspended matter.The initial two EOF modes were well correlated with the total suspended matter and back-scattering coefficient.Furthermore,the first EOF modes of the four classes of Rrs(A-D Rrs-EOF1)significantly contributed to the total variances of each Rrs class.In addition,the correlation coefficients between the amplitude factors of class A-D Rrs-EOF1 and the variances of the relevant water quality and optical parameters were better than those of the unclassified ones.The spectral shape of class ARrs-EOF1 was governed by the absorption characteristic of chlorophyll a and colored dissolved organic matter(CDOM).The spectral shape of class B Rrs-EOF1 was governed by the absorption characteristic of CDOM since it exhibited a high correlation with the absorption coefficient of CDOM(ag(λ)),whereas the spectral shape of class C Rrs-EOF1 was governed by the back-scattering characteristics but not affected by the suspended matter.The spectral shape of class D Rrs-EOF1 exhibited a relatively good correlation with all the water quality parameters,which played a significant role in deciding its spectral shape.
基金Project supported by the National Natural Science Foundation of China (No. 81271856)the National College of Innovative Experimental Projects of China (No. 101069819)
文摘To our knowledge,few reports on Demodex studied at the molecular level are available at present.In this study our group,for the first time,cloned,sequenced and analyzed the chitin synthase(CHS) gene fragments of Demodex folliculorum,Demodex brevis,and Demodex canis(three isolates from each species) from Xi'an China,by designing specific primers based on the only partial sequence of the CHS gene of D.canis from Japan,retrieved from GenBank.Results show that amplification was successful only in three D.canis isolates and one D.brevis isolate out of the nine Demodex isolates.The obtained fragments were sequenced to be 339 bp for D.canis and 338 bp for D.brevis.The CHS gene sequence similarities between the three Xi'an D.canis isolates and one Japanese D.canis isolate ranged from 99.7% to 100.0%,and those between four D.canis isolates and one D.brevis isolate were 99.1%-99.4%.Phylogenetic trees based on maximum parsimony(MP) and maximum likelihood(ML) methods shared the same clusters,according with the traditional classification.Two open reading frames(ORFs) were identified in each CHS gene sequenced,and their corresponding amino acid sequences were located at the catalytic domain.The relatively conserved sequences could be deduced to be a CHS class A gene,which is associated with chitin synthesis in the integument of Demodex mites.
文摘Despite controversy regarding the clinical value of semen analysis, male fertility investigation still relies on a standardized analysis of the semen parameters. This is especially true for infertility clinics in both developing and developed countries. Other optional tests or sophisticated technologies have not been widely applied. The current review addresses important changes in the analysis of semen as described in the new World Health Organization （WHO） manual for semen analysis. The most important change in the manual is the use of evidence-based publications as references to determine cutoff values for normality. Apart from the above mentioned changes, the initial evaluation and handling methods remain, in most instances, the same as in previous editions. Furthermore, the review evaluates the importance of quality control in andrology with emphasis on the evaluation of sperm morphology. WHO sperm morphology training programmes for Sub-Saharan countries were initiated at Tygerberg Hospital in 1995. The external qualitY control programme has ensured that the majority of participants have maintained their morphological reading skills acquired during initial training. This review reports on current sperm functional tests, such as the induced acrosome reaction, and sperm-zona pellucida binding assays, as well as the impact of sperm quality in terms of DNA integrity, and the relationship of sperm function tests to sperm morphology.
基金supported by grants from the National Natural Science Foundation of China(No.81670634)Graduate student scientific research innovation projects in Jiangsu province(No.KYLX15_0981)Nanjing Medical University Science and Technology Development Fund(No.2016NJMU065)
文摘Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.The gene expression profiles of CD11 b^(+)/Ly6 C^(intermediate)macrophages of the sham surgery mice,and the mice 4 h,24 h and 9 days after renal IRI were downloaded from the Gene Expression Omnibus database.Analysis of m RNA expression profiles was conducted to identify differentially expressed genes(DEGs),biological processes and pathways by the series test of cluster.Protein-protein interaction network was constructed and analysed to discover the key genes.A total of 6738 DEGs were identified and assigned to 20 model profiles.DEGs in profile 13 were one of the predominant expression profiles,which are involved in immune cell chemotaxis and proliferation.Signet analysis showed that Atp5 a1,Atp5 o,Cox4 i,Cdc42,Rac2 and Nhp2 were the key genes involved in oxidation-reduction,apoptosis,migration,M1-M2 differentiation,and proliferation of macrophages.RPS18 may be an appreciate reference gene as it was stable in macrophages.The identified DEGs and their enriched pathways investigate factors that may participate in the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.Moreover,the vital gene Nhp2 may involve the polarization of macrophages,which may be a new target to affect the process of AKI.
基金Supported by the National Natural Science Foundation of China (30870144)
文摘Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based on bioinformatic analysis, we found that Arabidopsis genome actually encoded 17 OVATE domain-containing proteins. One of them, AtOFP19, has not been previously identified. Based on their amino acid sequence similarity, AtOFPs proteins can be divided into two groups. Most of the AtOFPs were located in nuclear, four of them were presented in chloroplast and the remaining two members appeared in cytoplasmic. A genome- wide microarray based gene expression analysis involving 47 stages of vegetative and reproductive development revealed that AtOFPs have diverse expression pattems. Investigation of proteins interaction showed that nine AtOFPs only interacted with TALE homeodomain proteins, which are fundamental regulators of plant meristem function and leaf development. Our work could provide important leads toward functional genomics studies of ovate family proteins, which may be involved in a previously unrecognized control mechanism in plant development
基金supported by the National Natural Science Foundation of China(No.NSFC51608446)the Fundamental Research Fund for Central Universities of China(No.3102016ZY015)
文摘Traditional Global Sensitivity Analysis（GSA） focuses on ranking inputs according to their contributions to the output uncertainty.However,information about how the specific regions inside an input affect the output is beyond the traditional GSA techniques.To fully address this issue,in this work,two regional moment-independent importance measures,Regional Importance Measure based on Probability Density Function（RIMPDF） and Regional Importance Measure based on Cumulative Distribution Function（RIMCDF）,are introduced to find out the contributions of specific regions of an input to the whole output distribution.The two regional importance measures prove to be reasonable supplements of the traditional GSA techniques.The ideas of RIMPDF and RIMCDF are applied in two engineering examples to demonstrate that the regional moment-independent importance analysis can add more information concerning the contributions of model inputs.
基金supported by the Equipment Development Department ‘‘13th Five-year” Equipment Research Field Foundation of China Central Military Commission(No.6140244010216HT15001)
文摘Estimating the Probability Density Function（PDF） of the performance function is a direct way for structural reliability analysis,and the failure probability can be easily obtained by integration in the failure domain.However,efficiently estimating the PDF is still an urgent problem to be solved.The existing fractional moment based maximum entropy has provided a very advanced method for the PDF estimation,whereas the main shortcoming is that it limits the application of the reliability analysis method only to structures with independent inputs.While in fact,structures with correlated inputs always exist in engineering,thus this paper improves the maximum entropy method,and applies the Unscented Transformation（UT） technique to compute the fractional moments of the performance function for structures with correlations,which is a very efficient moment estimation method for models with any inputs.The proposed method can precisely estimate the probability distributions of performance functions for structures with correlations.Besides,the number of function evaluations of the proposed method in reliability analysis,which is determined by UT,is really small.Several examples are employed to illustrate the accuracy and advantages of the proposed method.
基金Supported by National Natural Science Foundation of China(Grant Nos.51135003,51205050,U1234208)Key National Science&Technology Special Project on"High-Grade CNC Machine Tools and Basic Manufacturing Equipments"(Grant No.2013ZX04011011)+1 种基金Research Fund for the Doctoral Program of Higher Education of China(Grant No.20110042120020)Fundamental Research Funds for the Central
文摘Reliability allocation of computerized numerical controlled(CNC)lathes is very important in industry.Traditional allocation methods only focus on high-failure rate components rather than moderate failure rate components,which is not applicable in some conditions.Aiming at solving the problem of CNC lathes reliability allocating,a comprehensive reliability allocation method based on cubic transformed functions of failure modes and effects analysis(FMEA)is presented.Firstly,conventional reliability allocation methods are introduced.Then the limitations of direct combination of comprehensive allocation method with the exponential transformed FMEA method are investigated.Subsequently,a cubic transformed function is established in order to overcome these limitations.Properties of the new transformed functions are discussed by considering the failure severity and the failure occurrence.Designers can choose appropriate transform amplitudes according to their requirements.Finally,a CNC lathe and a spindle system are used as an example to verify the new allocation method.Seven criteria are considered to compare the results of the new method with traditional methods.The allocation results indicate that the new method is more flexible than traditional methods.By employing the new cubic transformed function,the method covers a wider range of problems in CNC reliability allocation without losing the advantages of traditional methods.
文摘Statistical analysis was done on simultaneous wave and wind using data recorded by discus-shape wave buoy. The area is located in the southern Caspian Sea near the Anzali Port. Recorded wave data were obtained through directional spectrum wave analysis. Recorded wind direction and wind speed were obtained through the related time series as well. For 12-month measurements（May 25 2007-2008）, statistical calculations were done to specify the value of nonlinear auto-correlation of wave and wind using the probability distribution function of wave characteristics and statistical analysis in various time periods. The paper also presents and analyzes the amount of wave energy for the area mentioned on the basis of available database. Analyses showed a suitable comparison between the amounts of wave energy in different seasons. As a result, the best period for the largest amount of wave energy was known. Results showed that in the research period, the mean wave and wind auto correlation were about three hours. Among the probability distribution functions, i.e Weibull, Normal, Lognormal and Rayleigh, ＂Weibull＂ had the best consistency with experimental distribution function shown in different diagrams for each season. Results also showed that the mean wave energy in the research period was about 49.88 k W/m and the maximum density of wave energy was found in February and March, 2010.
基金supported by the Key Technology Research and Development Program of Shandong Province(No.2018GHY115007)。
文摘This study analyzed the function of different glutathione S-transferase(GST)isoforms and detoxification metabolism responses in Manila clam,Ruditapes philippinarum,exposed to 4 kinds of polycyclic aromatic hydrocarbons(PAHs)single,and their mixtures for 15 days under laboratory conditions.13 kinds of GSTs in R.philippinarum were classified,and the results of tissue distribution indicated that 12 kinds of GSTs(except GST sigma 3)expressed most in digestive glands.We detected the m RNA expression levels of aryl hydrocarbon receptor signaling pathway,and detoxification system in digestive glands of clams exposed to benzo[a]pyrene(Ba P),chrysene(CHR),benzo[a]anthracene(Ba A),benzo[b]fluoranthene(Bb F),and Ba P+CHR+Ba A+Bb F,respectively.Among these genes,we selected GST-sigma,GST-omega and GST-pi as potential indicators to Ba P;GST-sigma,GST-A and GST-rho to CHR;GST-pi,GST-sigma,GST-A,GST-rho and GST-microsomal to Ba A;GST-theta and GST-mu to Bb F;while GST-pi and GST-mu to the mixture of Ba P,CHR,Ba A and Bb F.Additionally,the bioaccumulation of PAHs in tissues increased remarkably over time,and showed an obvious doseeffect.Under the same concentration,the bioaccumulation in single exposure group was higher than that in mixture group,and the bioaccumulation of PAHs in tissues with different concentrations of stress was irregular.The results revealed the metabolic differences and bioaccumulation rules in clams exposed to four kinds of PAHs,and provided more valuable information for the PAHs risk assessment.
文摘In order to make rail transit smart stations serve passengers better,the potential of smart stations should be optimized to reduce time cost,and relieve traffic congestion.In this paper,the construction of smart stations based on the management experience of a subway station and the benefits of smart stations were analyzed and discussed.The construction of smart stations,as well as the key technologies for the construction of smart stations,will aid in the automation and intelligent management of subway stations.
基金supported by the National Natural Science Foundation of China(Grant No.32072049)Open Foundation of State Key Laboratory of Rice Biology,China(Grant No.20210208).
文摘An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.
