Genetic transformation is a crucial tool for investigating gene function and advancing molecular breeding in crops, with Agrobacterium tumefaciens-mediated transformation being the primary method for plant genetic mod...Genetic transformation is a crucial tool for investigating gene function and advancing molecular breeding in crops, with Agrobacterium tumefaciens-mediated transformation being the primary method for plant genetic modification. However, this approach exhibits significant genotypic dependence in maize. Therefore, to overcome these limitations, we combined dynamic transcriptome analysis and genome-wide association study (GWAS) to identify the key genes controlling Agrobacterium infection frequency (AIF) in immature maize embryos. Transcriptome analysis of Agrobacterium-infected embryos uncovered 8483 and 1580 genotype-specific response genes in the maize line 18-599R with low AIF and A188 with high AIF, respectively. A weighted gene co-expression network analysis (WGCNA) revealed five and seven stage-specific co-expression modules in each corresponding line. Based on a self-developed AIF quantitation method, the GWAS revealed 30 AIF-associated single-nucleotide polymorphisms and 315 candidate genes under multiple environments. Integration of GWAS and WGCNA further identified 12 key genes associated with high AIF in A188. ZmHRGP, encoding a hydroxyproline-rich glycoprotein, was functionally validated as a key factor of AIF in immature embryos. Knockout of ZmHRGP enabled us to establish a high-efficiency genetic transformation system for the 18-599R line, with the transformation frequency being approximately 80%. Moreover, the transient reduction of ZmHRGP expression significantly enhanced the AIF of maize calluses and leaves. Collectively, these findings advance our understanding of plant factors controlling Agrobacterium infection and contribute to developing more efficient Agrobacterium-mediated transformation systems in crops.展开更多
Host-pathogen interactions are highly complex,involving large dynamic changes in gene expression during infection. These interactions are fundamental to understanding anti-infection immunity of hosts, as well as the p...Host-pathogen interactions are highly complex,involving large dynamic changes in gene expression during infection. These interactions are fundamental to understanding anti-infection immunity of hosts, as well as the pathogenesis of pathogens. For bacterial pathogens interacting with animal hosts, timeresolved dual RNA-seq of infected tissue is difficult to perform due to low pathogen load in infected tissue. In this study, an acute infection model of Larimichthys crocea infected by Pseudomonas plecoglossicida was established. The spleens of infected fish exhibited typical symptoms, with a maximum bacterial load at two days post-injection(dpi). Time-resolved dual RNA-seq of infected spleens was successfully applied to study hostpathogen interactions between L. crocea and P.plecoglossicida. The spleens of infected L. crocea were subjected to dual RNA-seq, and transcriptome data were compared with those of noninfected spleens or in vitro cultured bacteria. Results showed that pathogen-host interactions were highly dynamically regulated, with corresponding fluctuations in host and pathogen transcriptomes during infection. The expression levels of many immunogenes involved in cytokine-cytokine receptor,Toll-like receptor signaling, and other immunerelated pathways were significantly up-regulated during the infection period. Furthermore, metabolic processes and the use of oxygen in L. crocea were strongly affected by P. plecoglossicida infection. The WGCNA results showed that the metabolic process was strongly related to the entire immune process.For P. plecoglossicida, the expression levels of motility-related genes and flagellum assemblyrelated genes were significantly up-regulated. The results of this study may help to elucidate the interactions between L. crocea and P.plecoglossicida.展开更多
Background:Aphis gossypii Glover(Hemiptera:Aphididae),a worldwide polyphagous phloem-feeding agricultural pest,has three wing morphs(winged parthenogenetic female,gynopara,and male)in the life cycle.The exclusive male...Background:Aphis gossypii Glover(Hemiptera:Aphididae),a worldwide polyphagous phloem-feeding agricultural pest,has three wing morphs(winged parthenogenetic female,gynopara,and male)in the life cycle.The exclusive males could fly from summer hosts to winter hosts,which are essential for gene exchanges of cotton aphid populations from different hosts or regions.However,the molecular mechanism of wing differentiation of male in A.gossypii remains unclear.Results:Morphological observation of male A.gossypii showed that there is no distinct difference in the external morphologies of the 1st and 2nd instar nymphs.The obvious differentiation of wing buds started in the 3rd instar nymph and was visible via naked eyes in the 4th instar nymphal stage,then adult male emerged with full wings.According to morphological dynamic changes,the development of wings in males were divided into four stages:preliminary stage(the 1st instar to 2nd instar),prophase(the 3rd instar),metaphase(the 4th instar),anaphase(the 5th instar).Results of feeding behavior monitoring via EPG(electrical penetration graph)technology indicated that although the male cotton aphids had strong desire to feed(longer duration of C 55.24%,F 5.05%and Pd waves 2.56%),its feeding efficiency to summer host cotton was low(shorter E13.56%and E2 waves 2.63%).Dynamic transcriptome analysis of male aphid at 5 different developmental periods showed that in the 3rd instar nymph,the number of up-regulated DEGs was significant increased,and time-course gene transcriptional pattern analyses results also showed that numerous genes categorized in clusters 3,5,and 8 had the highest expressed levels,which were consistent with morphological changes of wing buds.These results indicate that the 3rd instar nymph is the critical stage of wing bud differentiation in males.Furthermore,through pathway enrichment analysis of DEGs and WGCNA,it revealed that the neuroactive ligand-receptor interaction,Ras signaling pathway,dopaminergic synapse,circadian entrainment and the corresponding hub genes of PLK1,BUB1,SMC2,TUBG,ASPM,the kinesin family members(KIF23,KIF20,KIF18-19)and the novel subfamily of serine/threonine(Aurora kinase A and Aurora kinase B)probably played an important role in the critical stage of wing bud differentiation.Conclusion:This study explored morphological changes and genes transcriptional dynamics males in cotton aphid,revealed the phenomenon of low feeding efficiency of winged males on summer host cotton,and identified key signaling pathways and potential hub genes potentially involved in wing bud differentiation of male in A.gossypii.展开更多
S-dimethylarsino-glutathione(ZIO-101,darinaparsin®)exhibits broader antitumor activity and less toxicity than arsenic trioxide(ATO),a clinically used drug for acute promyelocytic leukemia(APL)treatment.However,th...S-dimethylarsino-glutathione(ZIO-101,darinaparsin®)exhibits broader antitumor activity and less toxicity than arsenic trioxide(ATO),a clinically used drug for acute promyelocytic leukemia(APL)treatment.However,the mechanisms of action underlying antileukemia activities of ZIO-101 remain largely unknown.Herein,by integrating dynamic transcriptomic analysis and biochemical characterizations,we for the first time delineated that ZIO-101 exerted antiproliferative effects against leukemia cells via activating ferroptosis pathway,a newly discovered iron-dependent programmed cell death,at the early stage,as evidenced by abnormally elevated intracellular iron contents and lipid peroxidation.We further demonstrated that silencing heme oxygenase 1(HMOX1),an important iron homeostasis-related gene,effectively attenuated ferroptosis induced by ZIO-101,with iron accumulation and lipid peroxidation being significantly alleviated.Significantly,we discovered that ZIO-101 and kinase inhibitors(Dasatinib/Dactolisib)could synergistically kill leukemia cells,with a combination index of<1.0 under all the tested drug concentrations.Our findings regarding the ferroptosis-mediated antileukemia activity of ZIO-101,based on the dynamic and temporal transcriptomic analysis,provide promising approaches to combat drug-resistant leukemia by combining ZIO-101 with kinase inhibitors.The methodology may be further exploited for uncovering the modes of action of other drugs.展开更多
基金supported by the National Key Research and Development Program of China(grant no.2021YFF1000303)the National Nature Science Foundation of China(grant nos.32072073 and 32372203)+2 种基金the Sichuan Province Science and Technology Support Program(grant no.2024NSFSC0313)the Shandong Provincial Natural Science Foundation(grant no.ZR2021QC098)the Biological Breeding Project of National Key Laboratory(grant no.SKL-ZD202211).
文摘Genetic transformation is a crucial tool for investigating gene function and advancing molecular breeding in crops, with Agrobacterium tumefaciens-mediated transformation being the primary method for plant genetic modification. However, this approach exhibits significant genotypic dependence in maize. Therefore, to overcome these limitations, we combined dynamic transcriptome analysis and genome-wide association study (GWAS) to identify the key genes controlling Agrobacterium infection frequency (AIF) in immature maize embryos. Transcriptome analysis of Agrobacterium-infected embryos uncovered 8483 and 1580 genotype-specific response genes in the maize line 18-599R with low AIF and A188 with high AIF, respectively. A weighted gene co-expression network analysis (WGCNA) revealed five and seven stage-specific co-expression modules in each corresponding line. Based on a self-developed AIF quantitation method, the GWAS revealed 30 AIF-associated single-nucleotide polymorphisms and 315 candidate genes under multiple environments. Integration of GWAS and WGCNA further identified 12 key genes associated with high AIF in A188. ZmHRGP, encoding a hydroxyproline-rich glycoprotein, was functionally validated as a key factor of AIF in immature embryos. Knockout of ZmHRGP enabled us to establish a high-efficiency genetic transformation system for the 18-599R line, with the transformation frequency being approximately 80%. Moreover, the transient reduction of ZmHRGP expression significantly enhanced the AIF of maize calluses and leaves. Collectively, these findings advance our understanding of plant factors controlling Agrobacterium infection and contribute to developing more efficient Agrobacterium-mediated transformation systems in crops.
基金the National Natural Science Foundation of China(31672694,31972836)Fujian Provincial Special Fund for Marine and Fishery Protection and Development(MCZ[2019]062)。
文摘Host-pathogen interactions are highly complex,involving large dynamic changes in gene expression during infection. These interactions are fundamental to understanding anti-infection immunity of hosts, as well as the pathogenesis of pathogens. For bacterial pathogens interacting with animal hosts, timeresolved dual RNA-seq of infected tissue is difficult to perform due to low pathogen load in infected tissue. In this study, an acute infection model of Larimichthys crocea infected by Pseudomonas plecoglossicida was established. The spleens of infected fish exhibited typical symptoms, with a maximum bacterial load at two days post-injection(dpi). Time-resolved dual RNA-seq of infected spleens was successfully applied to study hostpathogen interactions between L. crocea and P.plecoglossicida. The spleens of infected L. crocea were subjected to dual RNA-seq, and transcriptome data were compared with those of noninfected spleens or in vitro cultured bacteria. Results showed that pathogen-host interactions were highly dynamically regulated, with corresponding fluctuations in host and pathogen transcriptomes during infection. The expression levels of many immunogenes involved in cytokine-cytokine receptor,Toll-like receptor signaling, and other immunerelated pathways were significantly up-regulated during the infection period. Furthermore, metabolic processes and the use of oxygen in L. crocea were strongly affected by P. plecoglossicida infection. The WGCNA results showed that the metabolic process was strongly related to the entire immune process.For P. plecoglossicida, the expression levels of motility-related genes and flagellum assemblyrelated genes were significantly up-regulated. The results of this study may help to elucidate the interactions between L. crocea and P.plecoglossicida.
基金funded by National Natural Science Foundation of China(No.32102214)Special Fund for Basic Public Welfare Research of Institute of Cotton Research of CAAS(No.1610162020020604).
文摘Background:Aphis gossypii Glover(Hemiptera:Aphididae),a worldwide polyphagous phloem-feeding agricultural pest,has three wing morphs(winged parthenogenetic female,gynopara,and male)in the life cycle.The exclusive males could fly from summer hosts to winter hosts,which are essential for gene exchanges of cotton aphid populations from different hosts or regions.However,the molecular mechanism of wing differentiation of male in A.gossypii remains unclear.Results:Morphological observation of male A.gossypii showed that there is no distinct difference in the external morphologies of the 1st and 2nd instar nymphs.The obvious differentiation of wing buds started in the 3rd instar nymph and was visible via naked eyes in the 4th instar nymphal stage,then adult male emerged with full wings.According to morphological dynamic changes,the development of wings in males were divided into four stages:preliminary stage(the 1st instar to 2nd instar),prophase(the 3rd instar),metaphase(the 4th instar),anaphase(the 5th instar).Results of feeding behavior monitoring via EPG(electrical penetration graph)technology indicated that although the male cotton aphids had strong desire to feed(longer duration of C 55.24%,F 5.05%and Pd waves 2.56%),its feeding efficiency to summer host cotton was low(shorter E13.56%and E2 waves 2.63%).Dynamic transcriptome analysis of male aphid at 5 different developmental periods showed that in the 3rd instar nymph,the number of up-regulated DEGs was significant increased,and time-course gene transcriptional pattern analyses results also showed that numerous genes categorized in clusters 3,5,and 8 had the highest expressed levels,which were consistent with morphological changes of wing buds.These results indicate that the 3rd instar nymph is the critical stage of wing bud differentiation in males.Furthermore,through pathway enrichment analysis of DEGs and WGCNA,it revealed that the neuroactive ligand-receptor interaction,Ras signaling pathway,dopaminergic synapse,circadian entrainment and the corresponding hub genes of PLK1,BUB1,SMC2,TUBG,ASPM,the kinesin family members(KIF23,KIF20,KIF18-19)and the novel subfamily of serine/threonine(Aurora kinase A and Aurora kinase B)probably played an important role in the critical stage of wing bud differentiation.Conclusion:This study explored morphological changes and genes transcriptional dynamics males in cotton aphid,revealed the phenomenon of low feeding efficiency of winged males on summer host cotton,and identified key signaling pathways and potential hub genes potentially involved in wing bud differentiation of male in A.gossypii.
基金supported by the Research Grants Council of Hong Kong(nos.17307017 and R7070-18)the Innovation and Technology Commission of Hong Kong(no.ITS/124/17)the University of Hong Kong(for a scholarship to XHX).
文摘S-dimethylarsino-glutathione(ZIO-101,darinaparsin®)exhibits broader antitumor activity and less toxicity than arsenic trioxide(ATO),a clinically used drug for acute promyelocytic leukemia(APL)treatment.However,the mechanisms of action underlying antileukemia activities of ZIO-101 remain largely unknown.Herein,by integrating dynamic transcriptomic analysis and biochemical characterizations,we for the first time delineated that ZIO-101 exerted antiproliferative effects against leukemia cells via activating ferroptosis pathway,a newly discovered iron-dependent programmed cell death,at the early stage,as evidenced by abnormally elevated intracellular iron contents and lipid peroxidation.We further demonstrated that silencing heme oxygenase 1(HMOX1),an important iron homeostasis-related gene,effectively attenuated ferroptosis induced by ZIO-101,with iron accumulation and lipid peroxidation being significantly alleviated.Significantly,we discovered that ZIO-101 and kinase inhibitors(Dasatinib/Dactolisib)could synergistically kill leukemia cells,with a combination index of<1.0 under all the tested drug concentrations.Our findings regarding the ferroptosis-mediated antileukemia activity of ZIO-101,based on the dynamic and temporal transcriptomic analysis,provide promising approaches to combat drug-resistant leukemia by combining ZIO-101 with kinase inhibitors.The methodology may be further exploited for uncovering the modes of action of other drugs.
