Extreme environmental conditions such as temperature fluctuations,drought,and pathogen attacks can significantly impact plant growth,development,and productivity.Plants have evolved intricate enzymatic systems to miti...Extreme environmental conditions such as temperature fluctuations,drought,and pathogen attacks can significantly impact plant growth,development,and productivity.Plants have evolved intricate enzymatic systems to mitigate these stresses,among which GDSL esterase/lipase proteins (GELPs)-key members of the serine esterase/lipase superfamily-play important roles (Akoh et al.,2004).Characterized by a conserved GDSL motif and four essential amino acids (serine,glycine,asparagine,and histidine),GELPs exhibit versatile catalytic functions in lipid metabolism,cell wall modification,and stress responses (Ursache et al.,2021;Shen et al.,2022).展开更多
Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganod...Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganoderic acid(GA)biosynthesis of G.lucidum was limited.This study aimed to explore the functions of bHLH genes in ganoderic acid biosynthesis during G.lucidum growth development.Methods First,the genome-wide identification of GlbHLHs was performed through Hidden Markov model searches and Two-way blast.Furthermore,through physicochemical properties,gene structure,and phylogenetic analysis,as well as combining the transcriptome and metabolome data from different developmental stages of G.lucidum,candidate GlbHLHs were screened.Subsequently,their regulatory roles in ganoderic acid biosynthesis were explored using yeast one-hybrid and dual-luciferase reporter assays.Results A total of 11 GlbHLH members were characterized in G.lucidum.The upstream promoter regions of these genes enriched hormones and abiotic stress responsive elements.Although individual ganoderic acid monomers demonstrated marked differences in accumulation patterns across specific growth phases and tissue types,overall,the total GA content was consistently higher in caps than in stipes throughout development.In addition,all GlbHLHs exhibited high expression in whole G.lucidum from the primordium to maturation stages.Among them,GlbHLH5 and GlbHLH7 showed the highest expression in any stage and highly correlated with key genes associated with GA pathway.Functional validation through dual-luciferase assays and yeast one-hybrid experiments had demonstrated that GlbHLH5 activated the P2 region of the lanosterol synthase promoter,while GlbHLH7 activated the promoters of squalene epoxidase and squalene synthase.Conclusion Compared to plants,G.lucidum harbored a small number of bHLH members but all high expression in any stages.Additionally,GlbHLH5 and GlbHLH7 with the highest expression among GlbHLHs showed activation in regulating the biosynthesis of GA.These results provide a theoretical reference for further research on ganoderic acid regulation in G.lucidum,and thereby providing a molecular foundation for enhancing ganoderic acid yield to optimize the medicinal value of G.lucidum.展开更多
The fatty acid elongase 1(FAE1)genes of Brassic napus were cloned from two cultivars,i.e.Zhong-shuan No.9 with low erucic acid content,and Zhongyou 821 with high erucic acid content,using the degenerate PCR primers.Th...The fatty acid elongase 1(FAE1)genes of Brassic napus were cloned from two cultivars,i.e.Zhong-shuan No.9 with low erucic acid content,and Zhongyou 821 with high erucic acid content,using the degenerate PCR primers.The sequence analysis showed that there was no intron within the FAE1 genes.The FAE1 genes from Zhongyou 821 contained a coding sequence of 1521 nucleotides,and those cloned from Zhongshuan No.9 contained a 1517 bp coding sequence.Alignment of the FAE1 sequences from Brassica rapa,B.oleracea and B.napus detected 31 single nucleotide polymorphic sites(2.03%),which resulted in 7 amino-acid substitutions.Further analysis indicated that 19 SNPs were genome-specific,of which,95%were synonymous mutations.The nucleotide substitution at po-sition 1217 in the FAE1 genes led to a specific site of restricted cleavage.An AvrII cleavage site was present only in the C genome genes and absent in the A genome FAE1 genes.Digestion profile of the FAE1 sequences from B.rapa,B.oleracea and B.napus produced with AvrII confirmed that the FAE1 genes of B.oleracea origin was recognized and digested,while that of B.rapa origin could not.The results indicated that by AvrII cleavage it was possible to distinguish B.rapa from B.oleracea and be-tween the A and C genome of B.napus.In addition,the FAE1 genes could be used as marker genes to detect the pollen flow of B.napus,thus providing an alternative method for risk assessment of gene flow.展开更多
In the Kingdom of Saudi Arabia(KSA),thousands of plants are considered to have therapeutic value.The ambiguous use of identification mainly morphological characteristics of many plants has resulted in the adulteration...In the Kingdom of Saudi Arabia(KSA),thousands of plants are considered to have therapeutic value.The ambiguous use of identification mainly morphological characteristics of many plants has resulted in the adulteration and displacement of plant products which undermine their therapeutic value and weak documentation of plant resources.The aims of this study were therefore to evaluate genetic variability and explore the phylogeographic architecture for Saudi medicinal plant samples using rbcL and matK genes as barcodes for genomic identification.The matK and rbcL sequences collected for these samples were used as key markers for examining the relationship between Saudi medicinal plant species based on genetic diversity.During our study we were successful in identifying and documenting 4 different species(Foeniculum vulgare,Nitraria retusa,Dodonaea viscosa,and Rumex nervosus)located in Saudi Arabia using DNA barcoding technique.A total number of 8 sequences were obtained with a total sequence length of 6176 bp,where it ranged from 617 bp to 878 bp with an aver-age length of 772 bp.The total number of rbcL sequences length is 2801 bp,where it ranges from 617 bp to 807 bp with an average length of 700.2 bp.Out of the 4 plant samples used,only three samples were identified correctly on the species level with an identity percentage higher than 95%using rbcL gene.Additionally,4 matK sequences have been retrieved belong to 4 species.The total number of matK sequences length is 3375 bp,where it ranges from 819 bp to 878 bp with an average length of 843.8 bp.Out of the 4 plant samples used,only two samples were identified correctly on the species level with an identity percentage higher than 98% using matK gene.Both rbcL and matK have been able to identify most of our collected plant samples by genus,and some by species.Using only one DNA-barcoding technique was not reliable for plant identification,where matK and rbcL must be used as a dual DNA-barcoding procedure.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.32402580)the Science and Technology Program of Guangdong Province(Grant No.2023A0505090005)Modern Seed Industry Innovation Capability Enhancement Project of Guangdong Academy of Agricultural Sciences.
文摘Extreme environmental conditions such as temperature fluctuations,drought,and pathogen attacks can significantly impact plant growth,development,and productivity.Plants have evolved intricate enzymatic systems to mitigate these stresses,among which GDSL esterase/lipase proteins (GELPs)-key members of the serine esterase/lipase superfamily-play important roles (Akoh et al.,2004).Characterized by a conserved GDSL motif and four essential amino acids (serine,glycine,asparagine,and histidine),GELPs exhibit versatile catalytic functions in lipid metabolism,cell wall modification,and stress responses (Ursache et al.,2021;Shen et al.,2022).
基金funding from the Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(No.CI2021A04008)National Key Research and Development Project(No.2023YFC3504104)+2 种基金Hangzhou Joint Fund of the Zhejiang Provincial Natural Science Foundation of China(No.LHZSZ24H280003)Technology Major Program on Agricultural New Variety Breeding(No.2021C02073)Central Guiding Local Science and Technology Development Fund Project(No.2024ZY01009).
文摘Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganoderic acid(GA)biosynthesis of G.lucidum was limited.This study aimed to explore the functions of bHLH genes in ganoderic acid biosynthesis during G.lucidum growth development.Methods First,the genome-wide identification of GlbHLHs was performed through Hidden Markov model searches and Two-way blast.Furthermore,through physicochemical properties,gene structure,and phylogenetic analysis,as well as combining the transcriptome and metabolome data from different developmental stages of G.lucidum,candidate GlbHLHs were screened.Subsequently,their regulatory roles in ganoderic acid biosynthesis were explored using yeast one-hybrid and dual-luciferase reporter assays.Results A total of 11 GlbHLH members were characterized in G.lucidum.The upstream promoter regions of these genes enriched hormones and abiotic stress responsive elements.Although individual ganoderic acid monomers demonstrated marked differences in accumulation patterns across specific growth phases and tissue types,overall,the total GA content was consistently higher in caps than in stipes throughout development.In addition,all GlbHLHs exhibited high expression in whole G.lucidum from the primordium to maturation stages.Among them,GlbHLH5 and GlbHLH7 showed the highest expression in any stage and highly correlated with key genes associated with GA pathway.Functional validation through dual-luciferase assays and yeast one-hybrid experiments had demonstrated that GlbHLH5 activated the P2 region of the lanosterol synthase promoter,while GlbHLH7 activated the promoters of squalene epoxidase and squalene synthase.Conclusion Compared to plants,G.lucidum harbored a small number of bHLH members but all high expression in any stages.Additionally,GlbHLH5 and GlbHLH7 with the highest expression among GlbHLHs showed activation in regulating the biosynthesis of GA.These results provide a theoretical reference for further research on ganoderic acid regulation in G.lucidum,and thereby providing a molecular foundation for enhancing ganoderic acid yield to optimize the medicinal value of G.lucidum.
基金the National Natural Science Foundation of China(Grant No.30471099)Development Plan of the State Key Fundamental Research of China(Grant No.2006CB101600)the National High Technology and Development Program of China(Grant No.2006AA10A113)
文摘The fatty acid elongase 1(FAE1)genes of Brassic napus were cloned from two cultivars,i.e.Zhong-shuan No.9 with low erucic acid content,and Zhongyou 821 with high erucic acid content,using the degenerate PCR primers.The sequence analysis showed that there was no intron within the FAE1 genes.The FAE1 genes from Zhongyou 821 contained a coding sequence of 1521 nucleotides,and those cloned from Zhongshuan No.9 contained a 1517 bp coding sequence.Alignment of the FAE1 sequences from Brassica rapa,B.oleracea and B.napus detected 31 single nucleotide polymorphic sites(2.03%),which resulted in 7 amino-acid substitutions.Further analysis indicated that 19 SNPs were genome-specific,of which,95%were synonymous mutations.The nucleotide substitution at po-sition 1217 in the FAE1 genes led to a specific site of restricted cleavage.An AvrII cleavage site was present only in the C genome genes and absent in the A genome FAE1 genes.Digestion profile of the FAE1 sequences from B.rapa,B.oleracea and B.napus produced with AvrII confirmed that the FAE1 genes of B.oleracea origin was recognized and digested,while that of B.rapa origin could not.The results indicated that by AvrII cleavage it was possible to distinguish B.rapa from B.oleracea and be-tween the A and C genome of B.napus.In addition,the FAE1 genes could be used as marker genes to detect the pollen flow of B.napus,thus providing an alternative method for risk assessment of gene flow.
文摘In the Kingdom of Saudi Arabia(KSA),thousands of plants are considered to have therapeutic value.The ambiguous use of identification mainly morphological characteristics of many plants has resulted in the adulteration and displacement of plant products which undermine their therapeutic value and weak documentation of plant resources.The aims of this study were therefore to evaluate genetic variability and explore the phylogeographic architecture for Saudi medicinal plant samples using rbcL and matK genes as barcodes for genomic identification.The matK and rbcL sequences collected for these samples were used as key markers for examining the relationship between Saudi medicinal plant species based on genetic diversity.During our study we were successful in identifying and documenting 4 different species(Foeniculum vulgare,Nitraria retusa,Dodonaea viscosa,and Rumex nervosus)located in Saudi Arabia using DNA barcoding technique.A total number of 8 sequences were obtained with a total sequence length of 6176 bp,where it ranged from 617 bp to 878 bp with an aver-age length of 772 bp.The total number of rbcL sequences length is 2801 bp,where it ranges from 617 bp to 807 bp with an average length of 700.2 bp.Out of the 4 plant samples used,only three samples were identified correctly on the species level with an identity percentage higher than 95%using rbcL gene.Additionally,4 matK sequences have been retrieved belong to 4 species.The total number of matK sequences length is 3375 bp,where it ranges from 819 bp to 878 bp with an average length of 843.8 bp.Out of the 4 plant samples used,only two samples were identified correctly on the species level with an identity percentage higher than 98% using matK gene.Both rbcL and matK have been able to identify most of our collected plant samples by genus,and some by species.Using only one DNA-barcoding technique was not reliable for plant identification,where matK and rbcL must be used as a dual DNA-barcoding procedure.
