Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental st...Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.展开更多
Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-E...Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-Exp1 and Lc-Exp2 , were cloned. Lc-Exp1 and Lc-Exp2 was respectively composed of 531 bp encoding 177 amino acids and 537 bp encoding 179 amino acids. Eight cysteine residues and three tryp-tophan residues, which is supposed to be the characteristics of expansins, are conserved in both Lc-Exp1 and c-Exp2. In addition, the homology between the two expansins is 71. 6% at nucleotide acid sequences and 76.3% at amino acid sequences. The homology of Lc-Exp1 with Fa-Exp2 or Pp-Exp1 was 92.7% or 92.1%, but that of Lc-Exp2 with Fa-Exp2 or Pp-Exp1 was only 77. 4% or 76.3% at amino acid sequences.展开更多
Cucumber is one of the most widely consumed vegetables worldwide,and the fruit spine is an important fruit quality trait.Expansins play critical roles in fruit development;however,the regulation of expansins in cucumb...Cucumber is one of the most widely consumed vegetables worldwide,and the fruit spine is an important fruit quality trait.Expansins play critical roles in fruit development;however,the regulation of expansins in cucumber fruit spine development has not been reported.In this study,33 expansin genes were identified in the cucumber genome V3;additionally,expansin genes in Citrullus lanatus,Cucumis melo,Cucurbita maxima,Lagenaria siceraria,and Benincasa hispida were also identified.Phylogenetic analysis of expansin proteins in Cucurbitaceae and Arabidopsis showed that they evolved separately in each plant species.Phylogenetic analysis showed that C.maxima was derived earlier than the other five Cucurbitaceae species.The expression of CsEXPA2,CsEXPA14,and CsEXLA3 varied in cucumber lines with different fruit spine densities.A yeast two-hybrid assay showed that a putative auxin transporter encoded by numerous spine gene(ns)interacts with CsEXLA2,which may be involved in the development of the numerous spines in cucumber.These results provide novel insights into the expansins related to plant development and fruit spine development in cucumber.展开更多
Fruit firmness is an important quality trait of apple fruit texture,and the pre-harvest ripening period is the key period for the formation of apple fruit texture.Expansin is a cell wall loosing protein family that ha...Fruit firmness is an important quality trait of apple fruit texture,and the pre-harvest ripening period is the key period for the formation of apple fruit texture.Expansin is a cell wall loosing protein family that has four subfamilies:α-expansin(EXPA),β-expansin(EXPB),expansin-like A(EXLA),and expansin-like B(EXLB).In this study,we investigated the key period of pre-harvest texture formation in‘Golden Delicious’apples based on fruit longitudinal diameter,transverse diameter,firmness,tissue structure,respiration intensity,ethylene release rate,and expansin activity.Within the 10 days before harvest,the fruit was found to reach maturity.Semi-quantitative RT-PCR revealed that most of the expansins were expressed at the ripening stage before harvest.The biological function of the EXLB subfamily gene,MdEXLB1,was further identified,and its subcellular localization on the cell wall was confirmed by transient transformation experiments.Compared with the wild type(WT),the transgenic tomato lines overexpressing MdEXLB1 had lower plant height,earlier fruiting period,fewer days for fruit ripening,higher fruit maturity,lower fruit firmness,higher fruit expansin activity,more discrete flesh cell structure,and accelerated fruit ripening process.Overall,this is the first study to propose that the apple EXLB subfamily gene,MdEXLB1,has biological functions and plays an important role in promoting fruit ripening and softening.展开更多
Fruit ripening is a complex process and is regulated by many factors. Ethylene and polygalacturonase （PG）, lipoxygenase （LOX）, expansin （EXP） are all critical regulating factors in fruit ripening and softening p...Fruit ripening is a complex process and is regulated by many factors. Ethylene and polygalacturonase （PG）, lipoxygenase （LOX）, expansin （EXP） are all critical regulating factors in fruit ripening and softening process. With antisense ACS tomato, Nr mutant tomato and cultivated tomato as materials, Northern blot hybridization showed that PG, LeEXP1 and LOXexpressed differently in different parts of cultivated tomato fruit during ripening, which was related to fruit ripening. The ripening process of columella and radial pericarp was faster than pericarp. In both Nr mutant and antisense ACS transgenic tomato fruit, expression levels ofPG, LeEXPI and LOXwere generally lower than those in cultivated fruit but still related to fruit ripening. The expression levels ofPG, LeEXP1 and LOX increased in the mature green tomato fruits after 0.5 h treatment with ethylene （100 μL/L）. These results indicate that gene expression ofPG, LeEXP1 and LOXwere positively regulated by ethylene. The time and cumulative effect of the concentration exists in the expression of PG regulated by ethylene. The regulation of LOX expression mainly depended on the fruit development after great amount of ethylene was produced. PG played a major role in ripening and softening of tomato fruit, and cooperated with the regulation of EXP and LOX.展开更多
Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends ba...Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD （His-Phe-Asp） conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.展开更多
The introduction of reduced height （Rht） genes Rht-Blb and Rht-Dlb led to impressive increases in wheat （Triticum aestivum L.） yields during the Green Revolution. In the present study, the dynamic elongation of pe...The introduction of reduced height （Rht） genes Rht-Blb and Rht-Dlb led to impressive increases in wheat （Triticum aestivum L.） yields during the Green Revolution. In the present study, the dynamic elongation of peduncle in a set ofnear-isogenic lines （NILs） carrying different Rht alleles （Rht-Blb, Rht-Dlb, Rht-Blc, Rht-Dlb＋Rht-Blb, and Rht-Dlb＋Rht-Blc） were investigated. The reduction of the final length of peduncle in NILs was dependent mainly on the elongation rate, which was reduced by Rht genes, during rapid elongation phase. Resin sections showed that Rht genes strongly reduced the cell extension in peduncle. The expression of expansin genes, which mediate cell wall loosening and leading to cell expansion, were analysed by using real- time quantitative PCR （qPCR）. Among the 23 possible wheat expansin genes, 17 were expressed in the peduncle. The spatial distribution of expression was further analysed for five expansins that showed high expression levels in the peduncles of Rht lines. Compared to wild type plants, the incorporation of Rht-Dlb allele decreased about 37 and 80% of the expression levels of ExpA 7 and ExpA3 in elongation zone, respectively. The presence Rht-Blc dwarfing genes, however, produced 53% reduction in the expression level of ExpA7, and seriously decreased about 70% of ExpB9 expression. Although the expression levels of five genes exhibited variability among the lines, an expansin gene, ExpB2, showed its expression level highly associated with the cell elongation rate in peduncle of different Rht lines.展开更多
基金supported by the National Natural Science Foundation of China (Grant Nos.31902057 and 32072615)Zhejiang Provincial Natural Science Foundation of China (Grant No.LQ19C160012)the key research and development program of Zhejiang Province (Grant No.2021C02071)。
文摘Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.
文摘Using PCR degenerate primers, designed with reference to the sequences of the conserved amino acids of known expansins, to amplify cDNA fragments in litchi fruit by RT-PCR, two different cDNA fragments , named as Lc-Exp1 and Lc-Exp2 , were cloned. Lc-Exp1 and Lc-Exp2 was respectively composed of 531 bp encoding 177 amino acids and 537 bp encoding 179 amino acids. Eight cysteine residues and three tryp-tophan residues, which is supposed to be the characteristics of expansins, are conserved in both Lc-Exp1 and c-Exp2. In addition, the homology between the two expansins is 71. 6% at nucleotide acid sequences and 76.3% at amino acid sequences. The homology of Lc-Exp1 with Fa-Exp2 or Pp-Exp1 was 92.7% or 92.1%, but that of Lc-Exp2 with Fa-Exp2 or Pp-Exp1 was only 77. 4% or 76.3% at amino acid sequences.
基金supported by the earmarked fund for Modern Agro-industry Technology Research System (CARS-23)the National Natural Science Foundation of China (31672172)+1 种基金The Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-IVFCAAS)the Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture and Rural Affairs,China。
文摘Cucumber is one of the most widely consumed vegetables worldwide,and the fruit spine is an important fruit quality trait.Expansins play critical roles in fruit development;however,the regulation of expansins in cucumber fruit spine development has not been reported.In this study,33 expansin genes were identified in the cucumber genome V3;additionally,expansin genes in Citrullus lanatus,Cucumis melo,Cucurbita maxima,Lagenaria siceraria,and Benincasa hispida were also identified.Phylogenetic analysis of expansin proteins in Cucurbitaceae and Arabidopsis showed that they evolved separately in each plant species.Phylogenetic analysis showed that C.maxima was derived earlier than the other five Cucurbitaceae species.The expression of CsEXPA2,CsEXPA14,and CsEXLA3 varied in cucumber lines with different fruit spine densities.A yeast two-hybrid assay showed that a putative auxin transporter encoded by numerous spine gene(ns)interacts with CsEXLA2,which may be involved in the development of the numerous spines in cucumber.These results provide novel insights into the expansins related to plant development and fruit spine development in cucumber.
基金the National Key R&D Program of China(2016YFD0201100)the earmarked fund for China Agriculture Research System(CARS-27)+1 种基金the National Natural Science Foundation of China(31672116)the Natural Science Foundation of Hebei Province,China(C2021204134).
文摘Fruit firmness is an important quality trait of apple fruit texture,and the pre-harvest ripening period is the key period for the formation of apple fruit texture.Expansin is a cell wall loosing protein family that has four subfamilies:α-expansin(EXPA),β-expansin(EXPB),expansin-like A(EXLA),and expansin-like B(EXLB).In this study,we investigated the key period of pre-harvest texture formation in‘Golden Delicious’apples based on fruit longitudinal diameter,transverse diameter,firmness,tissue structure,respiration intensity,ethylene release rate,and expansin activity.Within the 10 days before harvest,the fruit was found to reach maturity.Semi-quantitative RT-PCR revealed that most of the expansins were expressed at the ripening stage before harvest.The biological function of the EXLB subfamily gene,MdEXLB1,was further identified,and its subcellular localization on the cell wall was confirmed by transient transformation experiments.Compared with the wild type(WT),the transgenic tomato lines overexpressing MdEXLB1 had lower plant height,earlier fruiting period,fewer days for fruit ripening,higher fruit maturity,lower fruit firmness,higher fruit expansin activity,more discrete flesh cell structure,and accelerated fruit ripening process.Overall,this is the first study to propose that the apple EXLB subfamily gene,MdEXLB1,has biological functions and plays an important role in promoting fruit ripening and softening.
基金Supported by National Project of Scientific and Technical Supporting Programs Funded by Ministry of Science and Technology of China (No.2006BAD22B01)National Natural Science Foundation of China (No.30800767)Postdoctoral Fund of China (No.20080430725)
文摘Fruit ripening is a complex process and is regulated by many factors. Ethylene and polygalacturonase （PG）, lipoxygenase （LOX）, expansin （EXP） are all critical regulating factors in fruit ripening and softening process. With antisense ACS tomato, Nr mutant tomato and cultivated tomato as materials, Northern blot hybridization showed that PG, LeEXP1 and LOXexpressed differently in different parts of cultivated tomato fruit during ripening, which was related to fruit ripening. The ripening process of columella and radial pericarp was faster than pericarp. In both Nr mutant and antisense ACS transgenic tomato fruit, expression levels ofPG, LeEXPI and LOXwere generally lower than those in cultivated fruit but still related to fruit ripening. The expression levels ofPG, LeEXP1 and LOX increased in the mature green tomato fruits after 0.5 h treatment with ethylene （100 μL/L）. These results indicate that gene expression ofPG, LeEXP1 and LOXwere positively regulated by ethylene. The time and cumulative effect of the concentration exists in the expression of PG regulated by ethylene. The regulation of LOX expression mainly depended on the fruit development after great amount of ethylene was produced. PG played a major role in ripening and softening of tomato fruit, and cooperated with the regulation of EXP and LOX.
基金supported by National Natural Science Foundation(Grant No.31270663)a grant from Agricultural Ministry of China(Grant No.2013ZX08009003-002-004)
文摘Expansins are cell-wall-loosening proteins that have multiple roles during plant development and stress- related processes. In this study, a novel expansin gene PwEXP2 was cloned by rapid amplification of cDNA ends based on the cDNA library of Picea wilsonii and EST fragment of PwEXP2. It was found that PwEXP2 coded 253 amino acids, and putative signal peptides exist at the N-terminal, followed by 8 cysteines, a HFD （His-Phe-Asp） conserved domain, and 4 tryptophan residues at the C-terminal. PwEXP2 was located in cytoplasm and nucleus when transformed in an onion epidermal cell. Quantitative real-time PCR assays showed that PwEXP2 was expressed in various tissues with a relatively high level in needles and low level in mature pollen. The expression level of PwEXP2 dramatically increased after seed germination. Gene expression profiles in abiotic stresses showed that PwEXP2 was induced by high temperature and osmotic stress but not involved in ABA-dependent signaling path- way. These results display the important roles of the PwEXP2 in plant development and multiple adversity stresses.
基金financed by the National High-Tech R&D Program of China (2011AA10A106)the Main Direction Program of Knowledge Innovation of Chinese Academy of Sciences (KSCX2-EW-N-02)+1 种基金the Promotive Research Fund for Excellent Young and Middle-Aged Scientists of Shandong Province, China (BS2013NY015)the Shandong Spark Program,China (2012XH11008)
文摘The introduction of reduced height （Rht） genes Rht-Blb and Rht-Dlb led to impressive increases in wheat （Triticum aestivum L.） yields during the Green Revolution. In the present study, the dynamic elongation of peduncle in a set ofnear-isogenic lines （NILs） carrying different Rht alleles （Rht-Blb, Rht-Dlb, Rht-Blc, Rht-Dlb＋Rht-Blb, and Rht-Dlb＋Rht-Blc） were investigated. The reduction of the final length of peduncle in NILs was dependent mainly on the elongation rate, which was reduced by Rht genes, during rapid elongation phase. Resin sections showed that Rht genes strongly reduced the cell extension in peduncle. The expression of expansin genes, which mediate cell wall loosening and leading to cell expansion, were analysed by using real- time quantitative PCR （qPCR）. Among the 23 possible wheat expansin genes, 17 were expressed in the peduncle. The spatial distribution of expression was further analysed for five expansins that showed high expression levels in the peduncles of Rht lines. Compared to wild type plants, the incorporation of Rht-Dlb allele decreased about 37 and 80% of the expression levels of ExpA 7 and ExpA3 in elongation zone, respectively. The presence Rht-Blc dwarfing genes, however, produced 53% reduction in the expression level of ExpA7, and seriously decreased about 70% of ExpB9 expression. Although the expression levels of five genes exhibited variability among the lines, an expansin gene, ExpB2, showed its expression level highly associated with the cell elongation rate in peduncle of different Rht lines.
