Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ...Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ascidians,including those expressed in the larval tail muscle,the adult body-wall muscle,and adult heart muscle.In this study,a novel striated non-tail muscle actin gene was identified from the RNA-seq data of Ciona savignyi embryos.Phylogenetic analysis,alignment of the N-terminal amino acid sequences and comparation of diagnostic residues provided evidence that it had high similarity with vertebrate cardiac and skeletal muscle actin.In situ hybridization and promoter-driven GFP reporter assay revealed that it was specifically expressed in the primordia of the oral and atrial siphon.We hereby defined it as siphon-specific muscle actin coding gene(Cs-SMA).A 201 bp(−1350 bp to−1150 bp)sequence containing T-box and Six1/2 binding motif within the upstream region of Cs-SMA confined the expression of GFP in the siphons of electroporated embryos.Six1/2 binding motif was experimentally confirmed to play indispensable role in controlling the siphon-specific expression of Cs-SMA.The tissue-specific expression of Cs-SMA in the siphon primordia indicated its potential crucial roles in Ciona embryogenesis and organogenesis.展开更多
Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtaine...Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.展开更多
A cDNA library containing 2×10￣6 recombinant plasmids was constructed using one week old young sorghum leaves as starting material and λgt10 as vector. In in situ hybridization, 2 positive clones were selected ...A cDNA library containing 2×10￣6 recombinant plasmids was constructed using one week old young sorghum leaves as starting material and λgt10 as vector. In in situ hybridization, 2 positive clones were selected using as probe RAcl cDNA from rice . One of them was subjected to Southern blot analysis and the recombinant containing the cDNA of SoAcl thus found was subcloned into pBluescript SK for sequencing. As shown by sequencing,SoAcl is composed of 1445 bps and 377 amino acids are encoded by the 1131 bp encoding region. It is the first report on it and has been included in the EMBO BANK (regist. no. X79378). High homology is found between it and other plant actin genes so far reported. The homology of amino acids sequency with RAcl is as high as 97.5%. Besides, highly homologous structural domains are found when it is compared with other plant and animal actins. These structural domains, thought to he sites for interaction among actins and/or between actins and their binding proteins, may prove to be important in the functioning of actin.(1 PhD candidate in Beijing University 2 Visiting Research Fellow of College of Oral Medicine of Beijing Medical University 3 Research Fellow of Institute of Developmental Biology, to whom correspondence is addressed 3 Reseach Fellow of Institute of Developmental Biology,to whom correspondence is addressed)展开更多
[Objectives]To develop methods for the early and rapid detection of tomato gray mold.[Methods]Utilizing the ACTIN gene of Botrytis cinerea as the target,a set of specific primers for loop-mediated isothermal amplifica...[Objectives]To develop methods for the early and rapid detection of tomato gray mold.[Methods]Utilizing the ACTIN gene of Botrytis cinerea as the target,a set of specific primers for loop-mediated isothermal amplification(LAMP)was designed and screened.Subsequently,the reaction system and conditions were optimized to achieve rapid isothermal amplification of B.cinerea.[Results]Through agarose gel electrophoresis and SYBR GreenⅠvisualization analysis,the optimal dosages of BstⅡDNA polymerase and dNTPs,as well as the optimal ratio of internal to external primers,were determined to be 0.6 U/μL,1.25 mmol/L,and 2:1,respectively.The specific detection of B.cinerea was accomplished at 61℃ for 40 min,achieving a sensitivity of 100 ag/μL,which is 106 times greater than that of conventional PCR detection.When this method was applied to the detection of tomato diseases,the detection limit for B.cinerea spores reached 20 spores/mL.Furthermore,the pathogen was detectable in tomato leaves that had been infected for 4 d,despite the absence of visible phenotypic symptoms of gray mold.[Conclusions]This method is suitable for the early,rapid,sensitive,and visual detection of tomato gray mold,thereby offering technical support for its early diagnosis,prevention,and control.展开更多
Cadmium(Cd)and Mercury(Hg)is among the heavy metals most hazardous for plant and human health.Known to induce oxidative stress in plants and disbalance equilibrium in the antioxidant defence system,these metals alter ...Cadmium(Cd)and Mercury(Hg)is among the heavy metals most hazardous for plant and human health.Known to induce oxidative stress in plants and disbalance equilibrium in the antioxidant defence system,these metals alter plant growth and cause damage at the cellular and molecular levels.Soybean is an important oilseed crop that is raised in soils often contaminated by Cd and Hg.The comparative studies on the deleterious effect of Cd and Hg and the defence system of antioxidants were not studied earlier in soybean plant.In this study,soybean plants were exposed to Cd(100μM CdCl_(2))and Hg(100μM HgCl_(2))and studied for physiological,biochemical and molecular responses.Both Cd and Hg treatment increased the magnitude of oxidative stress.Activities of antioxidant enzymes were significantly upregulated in response to Cd and Hg stress.Quantitative and qualitative assessment of isolated RNA showed significant differences in RNA under stress.Integrity values of RNA confirmed alterations.Transcript level of the Actin gene,involved in the morphogenesis of plants and also used as referenced gene in expression studies was analyzed using qRT-PCR just to check its stability and response under heavy metal stress.Results showed significant upregulation of the gene in the presence of Cd.It can be concluded that both Cd and Hg caused oxidative damage to plants,and adversely affected the quality of RNA.However,soybean tried to limit the adverse impacts of Cd and Hg stress by elevating the antioxidant system and upregulating Actin gene.展开更多
基金funded by the National Key Research and Development Program of China(Nos.2019YFE0190900,2018YFD0900705).
文摘Actin is a ubiquitous protein and plays essential roles on cellular structure maintenance and cellular motility in both muscle and non-muscle tissues.Multiple genes encoding muscle actin have been identified from the ascidians,including those expressed in the larval tail muscle,the adult body-wall muscle,and adult heart muscle.In this study,a novel striated non-tail muscle actin gene was identified from the RNA-seq data of Ciona savignyi embryos.Phylogenetic analysis,alignment of the N-terminal amino acid sequences and comparation of diagnostic residues provided evidence that it had high similarity with vertebrate cardiac and skeletal muscle actin.In situ hybridization and promoter-driven GFP reporter assay revealed that it was specifically expressed in the primordia of the oral and atrial siphon.We hereby defined it as siphon-specific muscle actin coding gene(Cs-SMA).A 201 bp(−1350 bp to−1150 bp)sequence containing T-box and Six1/2 binding motif within the upstream region of Cs-SMA confined the expression of GFP in the siphons of electroporated embryos.Six1/2 binding motif was experimentally confirmed to play indispensable role in controlling the siphon-specific expression of Cs-SMA.The tissue-specific expression of Cs-SMA in the siphon primordia indicated its potential crucial roles in Ciona embryogenesis and organogenesis.
基金Supported by the Postdoctoral Scientific Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘Degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the leaves of lris lacteal var.chinensis Fisch.Koidz. Actin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into pMD18-T vector. The positive clone identified by PCR was sequenced. The sequencing result showed that the Actin gene fragment from lris lacteal var.chinensis Fisch.Koidz contained about 598 bp, encoding 199 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank showed that it shared over 82% nueleotide sequence homology and 90% amino acid sequence homology. It indicated that this was the Actin gene. Because of the stability expression ofActin gene, it usually cited as the internal reference to study the expression and regulation of foundation in other genes of lris lacteal var.chinensis Fisch.Koidz well.
文摘A cDNA library containing 2×10￣6 recombinant plasmids was constructed using one week old young sorghum leaves as starting material and λgt10 as vector. In in situ hybridization, 2 positive clones were selected using as probe RAcl cDNA from rice . One of them was subjected to Southern blot analysis and the recombinant containing the cDNA of SoAcl thus found was subcloned into pBluescript SK for sequencing. As shown by sequencing,SoAcl is composed of 1445 bps and 377 amino acids are encoded by the 1131 bp encoding region. It is the first report on it and has been included in the EMBO BANK (regist. no. X79378). High homology is found between it and other plant actin genes so far reported. The homology of amino acids sequency with RAcl is as high as 97.5%. Besides, highly homologous structural domains are found when it is compared with other plant and animal actins. These structural domains, thought to he sites for interaction among actins and/or between actins and their binding proteins, may prove to be important in the functioning of actin.(1 PhD candidate in Beijing University 2 Visiting Research Fellow of College of Oral Medicine of Beijing Medical University 3 Research Fellow of Institute of Developmental Biology, to whom correspondence is addressed 3 Reseach Fellow of Institute of Developmental Biology,to whom correspondence is addressed)
基金Supported by Central Guiding Local Science and Technology Development Fund Project of Hebei Province(226Z6501G)Science and Technology Program of Hebei Academy of Sciences(23306,24306,25306).
文摘[Objectives]To develop methods for the early and rapid detection of tomato gray mold.[Methods]Utilizing the ACTIN gene of Botrytis cinerea as the target,a set of specific primers for loop-mediated isothermal amplification(LAMP)was designed and screened.Subsequently,the reaction system and conditions were optimized to achieve rapid isothermal amplification of B.cinerea.[Results]Through agarose gel electrophoresis and SYBR GreenⅠvisualization analysis,the optimal dosages of BstⅡDNA polymerase and dNTPs,as well as the optimal ratio of internal to external primers,were determined to be 0.6 U/μL,1.25 mmol/L,and 2:1,respectively.The specific detection of B.cinerea was accomplished at 61℃ for 40 min,achieving a sensitivity of 100 ag/μL,which is 106 times greater than that of conventional PCR detection.When this method was applied to the detection of tomato diseases,the detection limit for B.cinerea spores reached 20 spores/mL.Furthermore,the pathogen was detectable in tomato leaves that had been infected for 4 d,despite the absence of visible phenotypic symptoms of gray mold.[Conclusions]This method is suitable for the early,rapid,sensitive,and visual detection of tomato gray mold,thereby offering technical support for its early diagnosis,prevention,and control.
基金The authors would like to extend their sincere appreciation to the researchers supporting Project Number RSP2023R186,King Saud University,Riyadh,Saudi Arabia.
文摘Cadmium(Cd)and Mercury(Hg)is among the heavy metals most hazardous for plant and human health.Known to induce oxidative stress in plants and disbalance equilibrium in the antioxidant defence system,these metals alter plant growth and cause damage at the cellular and molecular levels.Soybean is an important oilseed crop that is raised in soils often contaminated by Cd and Hg.The comparative studies on the deleterious effect of Cd and Hg and the defence system of antioxidants were not studied earlier in soybean plant.In this study,soybean plants were exposed to Cd(100μM CdCl_(2))and Hg(100μM HgCl_(2))and studied for physiological,biochemical and molecular responses.Both Cd and Hg treatment increased the magnitude of oxidative stress.Activities of antioxidant enzymes were significantly upregulated in response to Cd and Hg stress.Quantitative and qualitative assessment of isolated RNA showed significant differences in RNA under stress.Integrity values of RNA confirmed alterations.Transcript level of the Actin gene,involved in the morphogenesis of plants and also used as referenced gene in expression studies was analyzed using qRT-PCR just to check its stability and response under heavy metal stress.Results showed significant upregulation of the gene in the presence of Cd.It can be concluded that both Cd and Hg caused oxidative damage to plants,and adversely affected the quality of RNA.However,soybean tried to limit the adverse impacts of Cd and Hg stress by elevating the antioxidant system and upregulating Actin gene.
