Kunitz protease inhibitors(KPIs)are ubiquitous in plants and act as crucial compounds in defense responses against insect attack and pathogen infection.However,the influence of gene duplication on the postdivergence o...Kunitz protease inhibitors(KPIs)are ubiquitous in plants and act as crucial compounds in defense responses against insect attack and pathogen infection.However,the influence of gene duplication on the postdivergence of the CsKPI genes involved in biotic stresses in tea plant is not well known.Here,we identified three CsKPI genes from tea plant(Camellia sinensis)and characterized their expression and evolutionary patterns among plant species.We found that CsKPI1,CsKPI2,and CsKPI3 diverged from their common ancestor 72.94 million years ago(MYA),and the tandem duplication of CsKPI2 and CsKPI3 occurred 26.78 MYA.An in vitro protein assay showed that the three CsKPI proteins were functional and inhibited the production of p-nitroanilide(PNA)from an artificial substrate.The three CsKPI-GFP fusion proteins localized to the cytoplasm.We showed that salicylic acid(SA)and transcripts of CsKPI2 and CsKPI3 significantly accumulated after infection with Glomerella cingulata.The application of exogenous SA stimulated the high expression of both CsKPI2 and CsKPI3 by activating cis-elements within their promoters.Under Ectropis oblique attack,CsKPI1 expression and jasmonic acid(JA)levels were more abundant in both insect-damaged leaf tissues and undamaged neighboring leaves.The application of jasmonic acid methyl ester elicited high expression levels of CsKPI1,suggesting that CsKPI1 accumulation requires JA production in tea plant.The overall findings suggest that the transcriptional divergence of KPI genes after duplication led to the specialized role of CsKPI1 in the physiological response to insect stress;the functional conservation between CsKPI2 and CsKPI3 confers resistance to pathogen infection in tea plant.展开更多
Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an i...Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an important phytohormone involving in plant stress.However,underlying molecular mechanisms of JA modulated osmotic stress response remains unclear.In this study,high concentration of mannitol induced JA accumulation and increase of peroxidase activity in tea plant.Integrated transcriptome mined a JA signaling master,MYC2 transcription factor is shown as a hub regulator that induced by mannitol,expression of which positively correlated with JA biosynthetic genes(LOX and AOS)and peroxidase genes(PER).CsMYC2 was determined as a nuclei-localized transcription activator,furthermore,ProteinDNA interaction analysis indicated that CsMYC2 was positive regulator that activated the transcription of CsLOX7,CsAOS2,CsPER1 and CsPER3via bound with their promoters,respectively.Suppression of CsMYC2 expression resulted in a reduced JA content and peroxidase activity and osmotic stress tolerance of tea plant.Overexpression of CsMYC2 in Arabidopsis improved JA content,peroxidase activity and plants tolerance against mannitol stress.Together,we proposed a positive feedback loop mediated by CsMYC2,CsLOX7 and CsAOS2 which constituted to increase the tolerance of osmotic stress through fine-tuning the accumulation of JA levels and increase of POD activity in tea plant.展开更多
Self-incompatibility(SI)is a kind of plant fertilization obstacle,which can prevent the harmful effects of inbreeding decline,but it hinders the breeding of inbred lines.Tea plants have SI and long-term cross-pollinat...Self-incompatibility(SI)is a kind of plant fertilization obstacle,which can prevent the harmful effects of inbreeding decline,but it hinders the breeding of inbred lines.Tea plants have SI and long-term cross-pollination,which limits the progress of genetic research and variety improvement.However,the mechanism of SI in tea plants is still a mystery.Herein,microscopic observation showed that the pollen tube could pass through the base of style and enter the ovary cavity after 48 h of self-pollination at different flowering stages,and the SI intensity at bud stage and full bloom stage was lower than initial bloom stage.RNA-seq analysis showed that 1,463 and 1,409 differentially expressed genes(DEGs)were associated with low SI at bud stage and full bloom stage,respectively,and 507 DEGs were associated with SI at initial bloom stage.The results of qRT-PCR validation of 20 DEGs were consistent with the RNA-seq data.Furthermore,CsRNS,CsSRKL5 and CsSRKL8 specifically expressed in style,which may be related to the low SI at bud stage,and three CsACC genes may be related to the low SI at full bloom stage.The results provide useful information for understanding the mechanism of SI in tea plants.展开更多
Tea processed from albino/etiolated tea shoots is favored by consumers because of its high theanine accumulation.To explore why theanine accumulates highly in new shoots of albino/etiolated tea cultivars,we compared t...Tea processed from albino/etiolated tea shoots is favored by consumers because of its high theanine accumulation.To explore why theanine accumulates highly in new shoots of albino/etiolated tea cultivars,we compared theanine content in shoots and roots of albino/etiolated and common green tea cultivars.Results suggested that high theanine accumulation in albino/etiolated tea shoots was likely not caused by higher theanine biosynthesis in roots.Further analyses suggested that CsAlaDC-catalyzed ethylamine biosynthesis and CsGOGAT1-catalyzed glutamate biosynthesis were more active,and CsGGT2-catalyzed theanine catabolism was weaker in new shoots of these albino/etiolated tea plant cultivars.Therefore,the high theanine accumulation in albino/etiolated shoots is probably contributed by the strong theanine biosynthesis and weak catabolism in new shoots.These findings provided more comprehensive insights into the high accumulation of theanine in new shoots of albino/etiolated tea cultivars,and the knowledge can be used in plant breeding for new cultivars with higher theanine accumulation.展开更多
Dear Editor,The tea plant(Camellia sinensis)is an evergreen species.Past breeding efforts have created purple leaf tea varieties with enhanced health benefits,such as‘Zijuan’(C.sinensis var.assa-mica cv.Zijuan[ZJ])(...Dear Editor,The tea plant(Camellia sinensis)is an evergreen species.Past breeding efforts have created purple leaf tea varieties with enhanced health benefits,such as‘Zijuan’(C.sinensis var.assa-mica cv.Zijuan[ZJ])(Figure 1A).To understand the mechanisms underlying the purple trait,the genome of ZJ was assembled using PacBio and Hi-C technologies(Figure 1B).The assembled genome size is approximately 3.06 Gb,comprising 1344 scaffolds with a scaffold N50 size of approximately 214.76 Mb(Supplemental Table 1).In addition,99.12%of the assembled sequences were anchored to 15 chromosomes.展开更多
Tea plant is an important economic crop,which is used to produce the world's oldest and most widely consumed tea beverages.Here,we present a high-quality reference genome assembly of the tea plant(Camellia sinensi...Tea plant is an important economic crop,which is used to produce the world's oldest and most widely consumed tea beverages.Here,we present a high-quality reference genome assembly of the tea plant(Camellia sinensis var.sinensis)consisting of 15 pseudo-chromosomes.LTR retrotransposons(LTR-RTs)account for 70.38%of the genome,and we present evidence that LTR-RTS play critical roles in genome size expansion and the transcriptional diversification of tea plant genes through preferential insertion in promoter regions and introns.Genes,particularly those coding for terpene biosynthesis pro-teins,associated with tea aroma and stress resistance were significantly amplified through recent tandem duplications and exist as gene clusters in tea plant genome.Phylogenetic analysis of the sequences of 81 tea plant accessions with diverse origins revealed three well-differentiated tea plant populations,support-ing the proposition for the southwest origin of the Chinese cultivated tea plant and its later spread to western Asia through introduction.Domestication and modern breeding left significant signatures on hundreds of genes in the tea plant genome,particularly those associated with tea quality and stress resis-tance.The genomic sequences of the reported reference and resequenced tea plant accessions provide valuable resources for future functional genomics study and molecular breeding of improved cul-tivars of tea plants.展开更多
基金supported by the National Key Research and Development Program of China(2018YFD1000601)the National Natural Science Foundation of China(31171608)+2 种基金the Special Innovative Province Construction in Anhui Province(15czs08032)the Special Project for Central Guiding Science and Technology Innovation of Region in Anhui Province(2016080503B024)the Program for Changjiang Scholars and Innovative Research Team in University(IRT1101).
文摘Kunitz protease inhibitors(KPIs)are ubiquitous in plants and act as crucial compounds in defense responses against insect attack and pathogen infection.However,the influence of gene duplication on the postdivergence of the CsKPI genes involved in biotic stresses in tea plant is not well known.Here,we identified three CsKPI genes from tea plant(Camellia sinensis)and characterized their expression and evolutionary patterns among plant species.We found that CsKPI1,CsKPI2,and CsKPI3 diverged from their common ancestor 72.94 million years ago(MYA),and the tandem duplication of CsKPI2 and CsKPI3 occurred 26.78 MYA.An in vitro protein assay showed that the three CsKPI proteins were functional and inhibited the production of p-nitroanilide(PNA)from an artificial substrate.The three CsKPI-GFP fusion proteins localized to the cytoplasm.We showed that salicylic acid(SA)and transcripts of CsKPI2 and CsKPI3 significantly accumulated after infection with Glomerella cingulata.The application of exogenous SA stimulated the high expression of both CsKPI2 and CsKPI3 by activating cis-elements within their promoters.Under Ectropis oblique attack,CsKPI1 expression and jasmonic acid(JA)levels were more abundant in both insect-damaged leaf tissues and undamaged neighboring leaves.The application of jasmonic acid methyl ester elicited high expression levels of CsKPI1,suggesting that CsKPI1 accumulation requires JA production in tea plant.The overall findings suggest that the transcriptional divergence of KPI genes after duplication led to the specialized role of CsKPI1 in the physiological response to insect stress;the functional conservation between CsKPI2 and CsKPI3 confers resistance to pathogen infection in tea plant.
基金supported by the National Natural Science Foundation of China(Grant Nos.32202542 and U20A2045)the Project of Major Science and Technology in Anhui Province(Grant No.202003a06020021)+2 种基金the Project of Science and Technology of Yunnan Province(Grant No.202102AE090038)Anhui Provincial Natural Science Foundation(Grant No.2108085QC121)the Natural Science Projects for Colleges and Universities in the Anhui Province(Grant No.KJ2021A0145)。
文摘Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an important phytohormone involving in plant stress.However,underlying molecular mechanisms of JA modulated osmotic stress response remains unclear.In this study,high concentration of mannitol induced JA accumulation and increase of peroxidase activity in tea plant.Integrated transcriptome mined a JA signaling master,MYC2 transcription factor is shown as a hub regulator that induced by mannitol,expression of which positively correlated with JA biosynthetic genes(LOX and AOS)and peroxidase genes(PER).CsMYC2 was determined as a nuclei-localized transcription activator,furthermore,ProteinDNA interaction analysis indicated that CsMYC2 was positive regulator that activated the transcription of CsLOX7,CsAOS2,CsPER1 and CsPER3via bound with their promoters,respectively.Suppression of CsMYC2 expression resulted in a reduced JA content and peroxidase activity and osmotic stress tolerance of tea plant.Overexpression of CsMYC2 in Arabidopsis improved JA content,peroxidase activity and plants tolerance against mannitol stress.Together,we proposed a positive feedback loop mediated by CsMYC2,CsLOX7 and CsAOS2 which constituted to increase the tolerance of osmotic stress through fine-tuning the accumulation of JA levels and increase of POD activity in tea plant.
基金supported by the Project of Major Science and Technology in Anhui Province(202003a06020021)the Project of Innovation and Training of Anhui Agricultural University(XJDC2021084)the National Natural Science Foundation of China(U20A2045).
文摘Self-incompatibility(SI)is a kind of plant fertilization obstacle,which can prevent the harmful effects of inbreeding decline,but it hinders the breeding of inbred lines.Tea plants have SI and long-term cross-pollination,which limits the progress of genetic research and variety improvement.However,the mechanism of SI in tea plants is still a mystery.Herein,microscopic observation showed that the pollen tube could pass through the base of style and enter the ovary cavity after 48 h of self-pollination at different flowering stages,and the SI intensity at bud stage and full bloom stage was lower than initial bloom stage.RNA-seq analysis showed that 1,463 and 1,409 differentially expressed genes(DEGs)were associated with low SI at bud stage and full bloom stage,respectively,and 507 DEGs were associated with SI at initial bloom stage.The results of qRT-PCR validation of 20 DEGs were consistent with the RNA-seq data.Furthermore,CsRNS,CsSRKL5 and CsSRKL8 specifically expressed in style,which may be related to the low SI at bud stage,and three CsACC genes may be related to the low SI at full bloom stage.The results provide useful information for understanding the mechanism of SI in tea plants.
基金supported by grants from the National Natural Science Foundation of China(32072624,U20A2045)the National Key R&D Program of China(2022YFF1003103,2021YFD1601101)the Base of Introducing Talents for Tea Plant Biology and Quality Chemistry(D20026).
文摘Tea processed from albino/etiolated tea shoots is favored by consumers because of its high theanine accumulation.To explore why theanine accumulates highly in new shoots of albino/etiolated tea cultivars,we compared theanine content in shoots and roots of albino/etiolated and common green tea cultivars.Results suggested that high theanine accumulation in albino/etiolated tea shoots was likely not caused by higher theanine biosynthesis in roots.Further analyses suggested that CsAlaDC-catalyzed ethylamine biosynthesis and CsGOGAT1-catalyzed glutamate biosynthesis were more active,and CsGGT2-catalyzed theanine catabolism was weaker in new shoots of these albino/etiolated tea plant cultivars.Therefore,the high theanine accumulation in albino/etiolated shoots is probably contributed by the strong theanine biosynthesis and weak catabolism in new shoots.These findings provided more comprehensive insights into the high accumulation of theanine in new shoots of albino/etiolated tea cultivars,and the knowledge can be used in plant breeding for new cultivars with higher theanine accumulation.
基金supported by the National Natural Science Foundation of China(U20A2045)the Base of Introducing Talents for Tea Plant Biology and Quality Chemistry(D20026)the Science and Technology Project of Yunnan Province(grant no.202102AE090038).
文摘Dear Editor,The tea plant(Camellia sinensis)is an evergreen species.Past breeding efforts have created purple leaf tea varieties with enhanced health benefits,such as‘Zijuan’(C.sinensis var.assa-mica cv.Zijuan[ZJ])(Figure 1A).To understand the mechanisms underlying the purple trait,the genome of ZJ was assembled using PacBio and Hi-C technologies(Figure 1B).The assembled genome size is approximately 3.06 Gb,comprising 1344 scaffolds with a scaffold N50 size of approximately 214.76 Mb(Supplemental Table 1).In addition,99.12%of the assembled sequences were anchored to 15 chromosomes.
基金This work was supported by the National Key Research and Development Program of China(2018YFD1000601 and 2019YFD1001601)the National Natural Science Foundation of China(31800180)+2 种基金the Natural Science Foundation of Anhui Province of China(1908085MC75)the China Postdoctoral Science Foundation(2017M621992)and the special funds for tea germplasm garden construction(2060502 and 201834040003).
文摘Tea plant is an important economic crop,which is used to produce the world's oldest and most widely consumed tea beverages.Here,we present a high-quality reference genome assembly of the tea plant(Camellia sinensis var.sinensis)consisting of 15 pseudo-chromosomes.LTR retrotransposons(LTR-RTs)account for 70.38%of the genome,and we present evidence that LTR-RTS play critical roles in genome size expansion and the transcriptional diversification of tea plant genes through preferential insertion in promoter regions and introns.Genes,particularly those coding for terpene biosynthesis pro-teins,associated with tea aroma and stress resistance were significantly amplified through recent tandem duplications and exist as gene clusters in tea plant genome.Phylogenetic analysis of the sequences of 81 tea plant accessions with diverse origins revealed three well-differentiated tea plant populations,support-ing the proposition for the southwest origin of the Chinese cultivated tea plant and its later spread to western Asia through introduction.Domestication and modern breeding left significant signatures on hundreds of genes in the tea plant genome,particularly those associated with tea quality and stress resis-tance.The genomic sequences of the reported reference and resequenced tea plant accessions provide valuable resources for future functional genomics study and molecular breeding of improved cul-tivars of tea plants.
