5-Aminolevulinic acid(ALA),is a novel plant growth regulator that can enhance plant tolerance against salt stress.However,the molecular mechanism of ALA is not well studied.In this study,ALA improved salt tolerance of...5-Aminolevulinic acid(ALA),is a novel plant growth regulator that can enhance plant tolerance against salt stress.However,the molecular mechanism of ALA is not well studied.In this study,ALA improved salt tolerance of apple(Malus×domestica'Gala')when the detached leaves or cultured calli were used as the materials.The expression of MdWRKY71,a WRKY transcription factor(TF)gene was found to be responsive to NaCl as well as ALA treatment.Functional analysis showed that overexpressing(OE)-MdWRKY71 significantly improved the salt tolerance of the transgenic apple,while RNA interfering(RNAi)-MdWRKY71 reduced the salt tolerance.However,exogenous ALA alleviated the salt damage in the RNAi-MdWRKY71 apple.When MdWRKY71 was transferred into tobacco,the salt tolerance of transgenic plants was enhanced,which was further improved by exogenous ALA.Subsequently,MdWRKY71 bound to the W-box of promoters of MdSOS2,MdNHX1,MdCLC-g,MdSOD1,MdCAT1 and MdAPX1,transcriptionally activating the gene expressions.Since the genes are responsible for Na+and Cl-transport and antioxidant enzyme activity respectively,it can be concluded that MdWRKY71,a new TF,is involved in ALA-improved salt tolerance by regulating ion homeostasis and redox homeostasis.These results provided new insights into the transcriptional regulatory mechanism of ALA in enhancing apple salt tolerance.展开更多
5-Aminolevulinic acid(ALA)is a novel plant growth regulator that has shown outstanding capability to promote stomatal opening.Starch degradation,catalyzed byβ-amylase(EC3.2.1.2,BAM),plays an important role in stomata...5-Aminolevulinic acid(ALA)is a novel plant growth regulator that has shown outstanding capability to promote stomatal opening.Starch degradation,catalyzed byβ-amylase(EC3.2.1.2,BAM),plays an important role in stomatal opening.However,whether the starch breakdown is involved in ALA-regulating stomatal movement is unclear.In the current study,we found that exogenous ALA effectively stimulated the starch breakdown in guard cells,increasedβ-amylase activity and promoted stomatal opening in leaves of apple(Malus×domestica).Based on genome-wide identification,we identified a total of 119 members of BAM gene family in ten commonly Rosaceae crops.Analyses of gene structure,motif identification,and gene pair collinearity revealed relative conservation among members within the same group or subgroup.Among these genes,MdBAM17 and other 12 genes were identified as the orthologous genes of AtBAM1,which is responsible for starch degradation to modulate the stomatal movement in Arabidopsis.qRT-PCR analysis revealed a positive correlation between the expressions of MdBAM17 and stomatal aperture,as well asβ-amylase activity,whereas a negative correlation was observed with the starch content.Subcellular localization analysis confirmed that MdBAM17 is a chloroplast protein,consistent with the AtBAM1.MdBAM17 was mainly expressed in guard cells and responsive to exogenous ALA.Overexpressing MdBAM17 increasedβ-amylase activity and promoted starch breakdown,leading to stomatal opening,which was further strengthened by ALA.RNA-interfering MdBAM17 decreasedβ-amylase activity,resulting in starch accumulation,and impairing the stomatal opening by ALA.However,modulation of MdBAM17 expression did not affect the levels of flavonols and H_(2)O_(2)in guard cells,suggesting that MdBAM17-promoted starch degradation may function at downstream of ROS signaling in the ALAregulated stomatal opening.Our findings provide new insights into the mechanisms of ALA-regulated stomatal movement.展开更多
Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and ...Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and heme molecules to assemble hemoglobin(Zhang et al., 2021). The erythroid-specific enzyme δ-aminolevulinate synthase 2 (ALAS2) is a key rate-limiting factor in heme biosynthesis,with its expression increasing in late-stage erythropoiesis to meet heme demands (Sadlon et al., 1999). Zebrafish (Danio rerio) is a well-established model for studying erythropoiesis due to its genetic tractability and optical transparency (Zhang and Hamza, 2019;Zhang et al., 2021). The Tol2-mediated Gal4-UAS system has been widely applied for gene and enhancer trapping in zebrafish (Asakawa and Kawakami, 2009). However, reliable Gal4 enhancer-trap lines for erythropoiesis remain limited. Here, we report a transgenic zebrafish line with erythroid-specific Gal4FF expression under the control of the endogenous alas2 promoter, offering a more precise erythroblast labeling than the gata1a reporter line. This model provides a valuable tool for erythroid-specific investigations of blood flow dynamics and gene function.展开更多
基金funded by the Natural Science Foundation of China(Grant Nos.32230097 and 32172512)the Jiangsu Agricultural Science and Technology Innovation Fund[Grant No.CX(20)2023]+1 种基金the Jiangsu Special Fund for Frontier Foundation Research of Carbon Peaking and Carbon Neutralization(Grant No.BK20220005)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions。
文摘5-Aminolevulinic acid(ALA),is a novel plant growth regulator that can enhance plant tolerance against salt stress.However,the molecular mechanism of ALA is not well studied.In this study,ALA improved salt tolerance of apple(Malus×domestica'Gala')when the detached leaves or cultured calli were used as the materials.The expression of MdWRKY71,a WRKY transcription factor(TF)gene was found to be responsive to NaCl as well as ALA treatment.Functional analysis showed that overexpressing(OE)-MdWRKY71 significantly improved the salt tolerance of the transgenic apple,while RNA interfering(RNAi)-MdWRKY71 reduced the salt tolerance.However,exogenous ALA alleviated the salt damage in the RNAi-MdWRKY71 apple.When MdWRKY71 was transferred into tobacco,the salt tolerance of transgenic plants was enhanced,which was further improved by exogenous ALA.Subsequently,MdWRKY71 bound to the W-box of promoters of MdSOS2,MdNHX1,MdCLC-g,MdSOD1,MdCAT1 and MdAPX1,transcriptionally activating the gene expressions.Since the genes are responsible for Na+and Cl-transport and antioxidant enzyme activity respectively,it can be concluded that MdWRKY71,a new TF,is involved in ALA-improved salt tolerance by regulating ion homeostasis and redox homeostasis.These results provided new insights into the transcriptional regulatory mechanism of ALA in enhancing apple salt tolerance.
基金supported by the Natural Science Foundation of China(Grant No.32172512)the Jiangsu Special Fund for Frontier Foundation Research of Carbon Peaking and Carbon Neutralization(Grant No.BK20220005)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions。
文摘5-Aminolevulinic acid(ALA)is a novel plant growth regulator that has shown outstanding capability to promote stomatal opening.Starch degradation,catalyzed byβ-amylase(EC3.2.1.2,BAM),plays an important role in stomatal opening.However,whether the starch breakdown is involved in ALA-regulating stomatal movement is unclear.In the current study,we found that exogenous ALA effectively stimulated the starch breakdown in guard cells,increasedβ-amylase activity and promoted stomatal opening in leaves of apple(Malus×domestica).Based on genome-wide identification,we identified a total of 119 members of BAM gene family in ten commonly Rosaceae crops.Analyses of gene structure,motif identification,and gene pair collinearity revealed relative conservation among members within the same group or subgroup.Among these genes,MdBAM17 and other 12 genes were identified as the orthologous genes of AtBAM1,which is responsible for starch degradation to modulate the stomatal movement in Arabidopsis.qRT-PCR analysis revealed a positive correlation between the expressions of MdBAM17 and stomatal aperture,as well asβ-amylase activity,whereas a negative correlation was observed with the starch content.Subcellular localization analysis confirmed that MdBAM17 is a chloroplast protein,consistent with the AtBAM1.MdBAM17 was mainly expressed in guard cells and responsive to exogenous ALA.Overexpressing MdBAM17 increasedβ-amylase activity and promoted starch breakdown,leading to stomatal opening,which was further strengthened by ALA.RNA-interfering MdBAM17 decreasedβ-amylase activity,resulting in starch accumulation,and impairing the stomatal opening by ALA.However,modulation of MdBAM17 expression did not affect the levels of flavonols and H_(2)O_(2)in guard cells,suggesting that MdBAM17-promoted starch degradation may function at downstream of ROS signaling in the ALAregulated stomatal opening.Our findings provide new insights into the mechanisms of ALA-regulated stomatal movement.
基金supported by the National Key Research and Development Plan of China (2023YFA1802000)the National Natural Science Foundation of China for Distinguished Young Scholars (31925014),the National Natural Science Foundation of China Key Program (32130033),the National Natural Science Foundation of Original Exploratory Program (32350006)+5 种基金the Key Research Program of Frontier Sciences,Chinese Academy of Sciences (ZDBS-LY-SM010)Shanghai Pilot Program for Basic Research-Chinese Academy of Sciences,Shanghai Branch (JCYJ-SHFY-2022-006)Shanghai Science Technology Innovation Action Plan for Basic Research Program (21JC1406300)Haihe laboratory of Cell Ecosystem Innovation Fund (24HHXBSS00011)CAS project for Young Scientists in Basic Research (YSBR-077)supported by Science and Technology Commission of Shanghai Municipality (Shanghai Rising-Star Program,23QA1411300)
文摘Erythroid cells, the predominant circulating blood cells, are essential for oxygen and carbon dioxide transport (Obeagu, 2024).Their production, erythropoiesis, involves the coordinated synthesis of globin chains and heme molecules to assemble hemoglobin(Zhang et al., 2021). The erythroid-specific enzyme δ-aminolevulinate synthase 2 (ALAS2) is a key rate-limiting factor in heme biosynthesis,with its expression increasing in late-stage erythropoiesis to meet heme demands (Sadlon et al., 1999). Zebrafish (Danio rerio) is a well-established model for studying erythropoiesis due to its genetic tractability and optical transparency (Zhang and Hamza, 2019;Zhang et al., 2021). The Tol2-mediated Gal4-UAS system has been widely applied for gene and enhancer trapping in zebrafish (Asakawa and Kawakami, 2009). However, reliable Gal4 enhancer-trap lines for erythropoiesis remain limited. Here, we report a transgenic zebrafish line with erythroid-specific Gal4FF expression under the control of the endogenous alas2 promoter, offering a more precise erythroblast labeling than the gata1a reporter line. This model provides a valuable tool for erythroid-specific investigations of blood flow dynamics and gene function.
