Pollination methods play an important role in developing F1 populations.Our study presents an improved method of petal-assisted artificial pollination in tea plants.The field operation using this method exhibited high...Pollination methods play an important role in developing F1 populations.Our study presents an improved method of petal-assisted artificial pollination in tea plants.The field operation using this method exhibited high efficiency in which more than 500 pistillate flowers per day can be pollinated per capita,and the pollen amount from only one staminate flower was sufficient to pollinate more than five pistillate flowers.Additionally,we surveyed the fruit-setting ratio in three F1 groups,which were established using this method and the influence of rainfall on the fruit-setting ratio.The results indicate that the fruit-setting ratio was superior among the tested F1 population with an average value of 12.90%,and a significant negative correlation was observed between rainfall and fruit-setting ratio.By analyzing the growth states of each F1 population,both a high germination rate and survival rate were observed.Furthermore,the purity for 184 individuals in the F1 population of'Jiaming 1'×'Longjing 43'(J×L)were analyzed with 32 selected simple sequence repeats(SSRs).The purposes of this paper are(1)to demonstrate that this method is beneficial to establish high purity F1 populations and(2)to identify the number of SSR markers that are suitable for identification.The parentage analysis detected nine false-hybrid individuals,and the rate of false-hybrid was only 4.89%at 95%confidence,demonstrating that a low false-hybrid rate can be achieved using the method.Furthermore,one to two SSR markers randomly selected from each linkage group or chromosome were recommended as a sufficient quantity for SSR identification.展开更多
Albino tea,a type of tea closely associated with the chlorophyll-deficient phenotype is of great interest due to its multiple benefits to human health.To explore the potential mechanisms involved in albino tea,we perf...Albino tea,a type of tea closely associated with the chlorophyll-deficient phenotype is of great interest due to its multiple benefits to human health.To explore the potential mechanisms involved in albino tea,we performed metabolomics,DNA methylation and RNA-Seq analysis of green and albino leaves in a special tea mutant'Haishun 2'.The albino leaves accumulated significantly less catechins compared with the green leaves,which is closely associated with their difference in chlorophyll formation.A total of 385 candidate genes were identified by the integrative analysis.Two NAC transcription factors were less methylated and highly up regulated in the albino leaves of'Haishun 2',which was the first identification of the key roles of NAC transcription factors in albino tea and worth further study.Many genes involved in catechin biosynthesis were down regulated in albino leaves,such as 4-coumarate-CoA ligase,flavonol synthase and leucoanthocyanidin reductase genes,suggesting chlorophyll formation is important for catechin biosynthesis.Furthermore,CsMYB5 and CsGSTF11 were found to be differentially methylated and expressed in albino and green leaves,indicating their potential roles in catechin accumulation.This was the first report of the integrative analysis of transcriptome and DNA methylation data in tea plants and the results suggest that the integrative analysis is useful for exploring key genes associated with epigenetic changes in tea plants.展开更多
Background:Tumor-associated neutrophils(TANs)play a critical role in modulating immune responses and exhibit significant heterogeneity.Our previous study demonstrated that jagged canonical Notch ligand 2(JAG2)+TANs we...Background:Tumor-associated neutrophils(TANs)play a critical role in modulating immune responses and exhibit significant heterogeneity.Our previous study demonstrated that jagged canonical Notch ligand 2(JAG2)+TANs were associated with an immunosuppressive microenvironment in high-grade serous ovarian cancer(HGSOC),but the underlying mechanism remains unclear.This study aimed to elucidate the role of JAG2^(+)TANs in tumor immunosuppressive microenvironment in HGSOC.Methods:HGSOC samples were collected,with 274 samples constituting two independent cohorts(training and validation cohorts)and an additional 30 samples utilized to establish patient-derived tumor organoids(PDTOs).We characterized the number and phenotype of JAG2^(+)TANs by multiplex immunohistochemistry,flow cytometry,and single-cell RNA sequencing(scRNA-seq).We investigated the biological functions of JAG2 in immune evasion using in vitro co-culture systems,flow cytometry,tumor-bearing mouse models,and PDTOs.Results:JAG2^(+)TANs expressed elevated levels of immunosuppressive molecules,including programmed cell death ligand 1 and CD14,and had independent prognostic value for the overall survival of patients with HGSOC.scRNA-seq analysis revealed that JAG2^(+)TANs exhibited a terminally mature phenotype.The infiltration of JAG2^(+)TANs was positively correlated with the abundance of effector regulatory T cells(eTregs).Interaction with JAG2^(+)TANs skewed CD4+T cells towards an eTreg phenotype,a process that was suppressed by the Notch inhibitor LY3039478 and induced by recombinant Jagged2.Furthermore,we demonstrated that JAG2^(+)TANs enhanced Notch signaling activation,ultimately promoting recombination signal binding protein for immunoglobulin kappa J region(RBPJ)-induced differentiation of naïve CD4+T cells into eTregs.Clinically,JAG2^(+)TANs could serve as a biomarker for assessing immunotherapy resistance in various solid tumors.Pharmacological targeting of Notch signaling with LY3039478 or JAG2 neutralization antibodies enhanced the efficacy of programmed cell death protein 1(PD-1)monoclonal antibodies(mAbs)in both xenograft and PDTO models.Conclusions:The emergence of JAG2^(+)TANs is crucial for the differentiation of eTregs,which triggers immune evasion and resistance to anti-PD-1 therapy.Inhibiting Notch signaling with LY3039478 or JAG2 neutralization antibodies may overcome this anti-PD-1 resistance in HGSOC.展开更多
基金supported by the Chinese Modern Agricultural Industrial Technological System(CARS-19)the Key Scientific and Technological Project for New Variety of Tea Plant Breeding of Zhejiang Province(2016C02053).
文摘Pollination methods play an important role in developing F1 populations.Our study presents an improved method of petal-assisted artificial pollination in tea plants.The field operation using this method exhibited high efficiency in which more than 500 pistillate flowers per day can be pollinated per capita,and the pollen amount from only one staminate flower was sufficient to pollinate more than five pistillate flowers.Additionally,we surveyed the fruit-setting ratio in three F1 groups,which were established using this method and the influence of rainfall on the fruit-setting ratio.The results indicate that the fruit-setting ratio was superior among the tested F1 population with an average value of 12.90%,and a significant negative correlation was observed between rainfall and fruit-setting ratio.By analyzing the growth states of each F1 population,both a high germination rate and survival rate were observed.Furthermore,the purity for 184 individuals in the F1 population of'Jiaming 1'×'Longjing 43'(J×L)were analyzed with 32 selected simple sequence repeats(SSRs).The purposes of this paper are(1)to demonstrate that this method is beneficial to establish high purity F1 populations and(2)to identify the number of SSR markers that are suitable for identification.The parentage analysis detected nine false-hybrid individuals,and the rate of false-hybrid was only 4.89%at 95%confidence,demonstrating that a low false-hybrid rate can be achieved using the method.Furthermore,one to two SSR markers randomly selected from each linkage group or chromosome were recommended as a sufficient quantity for SSR identification.
基金the Central Publicinterest Scientific Institution Basal Research Fund(1610212020002)the Major Project of Agricultural Science and Technology in Breeding of Tea Plant Variety in Zhejiang Province(2021C02067)Earmarked Fund for China Agriculture Research System(CARS-19).
文摘Albino tea,a type of tea closely associated with the chlorophyll-deficient phenotype is of great interest due to its multiple benefits to human health.To explore the potential mechanisms involved in albino tea,we performed metabolomics,DNA methylation and RNA-Seq analysis of green and albino leaves in a special tea mutant'Haishun 2'.The albino leaves accumulated significantly less catechins compared with the green leaves,which is closely associated with their difference in chlorophyll formation.A total of 385 candidate genes were identified by the integrative analysis.Two NAC transcription factors were less methylated and highly up regulated in the albino leaves of'Haishun 2',which was the first identification of the key roles of NAC transcription factors in albino tea and worth further study.Many genes involved in catechin biosynthesis were down regulated in albino leaves,such as 4-coumarate-CoA ligase,flavonol synthase and leucoanthocyanidin reductase genes,suggesting chlorophyll formation is important for catechin biosynthesis.Furthermore,CsMYB5 and CsGSTF11 were found to be differentially methylated and expressed in albino and green leaves,indicating their potential roles in catechin accumulation.This was the first report of the integrative analysis of transcriptome and DNA methylation data in tea plants and the results suggest that the integrative analysis is useful for exploring key genes associated with epigenetic changes in tea plants.
基金supported by the Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases,the Obstetrics and Gynecology Hospital,Fudan University,and Shanghai Cancer Center,Fudan Universityfunded by grants from the National Natural Science Foundation of China(82072881,82273205,82203665,82473274)+1 种基金Natural Science Foundation of Shanghai(23ZR1408300),Shanghai Science and Technology Innovation Action Plan(23Y11909500,23Y11901800)Shanghai Clinical Research Center for Gynecological Diseases(22MC1940200).
文摘Background:Tumor-associated neutrophils(TANs)play a critical role in modulating immune responses and exhibit significant heterogeneity.Our previous study demonstrated that jagged canonical Notch ligand 2(JAG2)+TANs were associated with an immunosuppressive microenvironment in high-grade serous ovarian cancer(HGSOC),but the underlying mechanism remains unclear.This study aimed to elucidate the role of JAG2^(+)TANs in tumor immunosuppressive microenvironment in HGSOC.Methods:HGSOC samples were collected,with 274 samples constituting two independent cohorts(training and validation cohorts)and an additional 30 samples utilized to establish patient-derived tumor organoids(PDTOs).We characterized the number and phenotype of JAG2^(+)TANs by multiplex immunohistochemistry,flow cytometry,and single-cell RNA sequencing(scRNA-seq).We investigated the biological functions of JAG2 in immune evasion using in vitro co-culture systems,flow cytometry,tumor-bearing mouse models,and PDTOs.Results:JAG2^(+)TANs expressed elevated levels of immunosuppressive molecules,including programmed cell death ligand 1 and CD14,and had independent prognostic value for the overall survival of patients with HGSOC.scRNA-seq analysis revealed that JAG2^(+)TANs exhibited a terminally mature phenotype.The infiltration of JAG2^(+)TANs was positively correlated with the abundance of effector regulatory T cells(eTregs).Interaction with JAG2^(+)TANs skewed CD4+T cells towards an eTreg phenotype,a process that was suppressed by the Notch inhibitor LY3039478 and induced by recombinant Jagged2.Furthermore,we demonstrated that JAG2^(+)TANs enhanced Notch signaling activation,ultimately promoting recombination signal binding protein for immunoglobulin kappa J region(RBPJ)-induced differentiation of naïve CD4+T cells into eTregs.Clinically,JAG2^(+)TANs could serve as a biomarker for assessing immunotherapy resistance in various solid tumors.Pharmacological targeting of Notch signaling with LY3039478 or JAG2 neutralization antibodies enhanced the efficacy of programmed cell death protein 1(PD-1)monoclonal antibodies(mAbs)in both xenograft and PDTO models.Conclusions:The emergence of JAG2^(+)TANs is crucial for the differentiation of eTregs,which triggers immune evasion and resistance to anti-PD-1 therapy.Inhibiting Notch signaling with LY3039478 or JAG2 neutralization antibodies may overcome this anti-PD-1 resistance in HGSOC.
