[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, wit...[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, with a total length of 1 457 bp, coding 440 amino acids. [Result] Phylogenetic analysis showed that Capunl was closest to Pun1 of C. chinense, with a genetic distance of 0.019 3. Plant expression vector pCAM-Punl-GFP was constructed and transformed into to- bacco, and it was found that the protein coded by fusion gene Punl：：GFP was lo- cated on cell membrane. Prokaryotic expression vectors were constructed, and by SDS-PAGE and Western Blot detection, an induced protein with a molecular weight of 63 ku was obtained. It was found by real-time fluorescence quantitative expres- sion that Pun1 gene was expressed at the highest level 30 d after flowering, de- creased then, and could not be detected substantially 40 and 45 d after flowering. [Conclusion] This study provides information and reference for molecular regulation mechanism of Pun1 gene.展开更多
An attempt was made in an effort to synthesize a series of capsaicinoids, most of which are synthesized in our laboratory and characterized to be completely new members of capsaicinoids. The Structure-pungency depende...An attempt was made in an effort to synthesize a series of capsaicinoids, most of which are synthesized in our laboratory and characterized to be completely new members of capsaicinoids. The Structure-pungency dependence are presented and discussed.展开更多
The participation of O-methyltransferase (COMT) in phenylpropanoid-mediated capsaicinoid biosynthesis has long been proposed. Ferulic acid, a phenylpropanoid intermediate, is a precursor of capsaicinoid biosynthesis a...The participation of O-methyltransferase (COMT) in phenylpropanoid-mediated capsaicinoid biosynthesis has long been proposed. Ferulic acid, a phenylpropanoid intermediate, is a precursor of capsaicinoid biosynthesis and is produced from caffeic acid by the action of COMT. As previously reported that silencing Comt expression caused a drastic decrease in capsaicinoid accumulation, it was presumed that a Comt loss-of-function mutation would cause loss of pungency in Capsicum. This hypothesis was tested by cloning Comt1 and Comt2 from the placenta tissue of the pungent cultivar Habanero. The phylogenetic analysis and comparison of critical amino-acid residues for enzyme function showed that the two COMTs had high similarity with the COMTs of other plant species. Moreover, as the two Comts were both expressed in placenta tissue and expressed prior to the accumulation of capsaicinoids, the two genes could be candidates for capsaicinoid biosynthesis. Second, Comt1 loss-of-function mutants were screened from the germplasm. A truncated Comt1 transcript was expressed in non-pungent pepper No.3341 caused by deletion of the genomic region. The predicted No.3341 COMT1 lacked His-265, which was absolutely necessary for enzymatic activity. Contrary to our expectations, the Comt1 mutation was not related to non-pungency of No.3341, as the deletion of Comt1 did not co-segregate with non-pungency in the F2 population obtained from crossing No.3341 with Habanero. This result was confirmed by screening several pungent accessions harboring the same Comt1 deletion mutation. Although the participation of COMT in phenylpropanoid-mediated capsaicinoid biosynthesis has long been proposed, our present study shows that Comt1 can not be a target for controlling fruit pungency.展开更多
Ten capsaicin analogues were synthesized and their pungency degrees were determined through Scoville Organoleptic Test. The relationship between the structure and pungency degree of these capsaicin analogues was discu...Ten capsaicin analogues were synthesized and their pungency degrees were determined through Scoville Organoleptic Test. The relationship between the structure and pungency degree of these capsaicin analogues was discussed. Then four of these capsaicin analogues with higher pungency degree were picked out and added to anti-biofouling paints as repellents to study their anti-biofouling performance by shallow sea buoyant raft hung-plate experimentation. The results showed that capsaicin and dihydrocapsaicin exhibited equally good anti-biofouling performance while nordihydrocapsaicin and N-vanillylnonanamide had poor anti-biofouling performance. Experimental results also showed that the paints with only 0.1% capsaicin or dihydrocapsaicin as repellent without any other biocides had also exhibited good anti-biofouling performance, which provided a new idea for developing novel, more environment-friendly and Cu20-free antifouling paints.展开更多
The transcriptional cascade and regulatory loop play crucial roles in regulating plant-specialized metabolite biosynthesis.Capsaicinoids are unique to the genus Capsicum and confer a pungent flavor to its fruits.Howev...The transcriptional cascade and regulatory loop play crucial roles in regulating plant-specialized metabolite biosynthesis.Capsaicinoids are unique to the genus Capsicum and confer a pungent flavor to its fruits.However,the transcriptional regulation of capsaicinoid biosynthesis remains largely unknown.In this study,two AP2/ERF transcription factors(TFs),CaERF102 and CaERF111,were characterized for their role in the capsaicinoid biosynthesis process.Expression analysis of two ERFs and capsaicinoid biosynthetic genes(CBGs)suggested that they were associated with capsaicinoid biosynthesis.Both ERFs encode nuclear-localized proteins and function as transcriptional activators through their C-terminal activation motifs.The two ERF TFs participated in capsaicinoid biosynthesis by directly activating the promoters of key CBGs,and this activation was significantly enhanced when CaMYC2 was co-expressed.Moreover,CaERF102 and CaERF111 were found to interact with CaMYC2.This study helps elucidate the AP2/ERF TF regulatory network that governs capsaicinoid biosynthesis in Capsicum species.展开更多
Pepper(Capsicum spp.)is an important vegetable crop that provides a unique pungent sensation when eaten.Through construction of a pepper variome map,we examined the main groups that emerged during domestication and br...Pepper(Capsicum spp.)is an important vegetable crop that provides a unique pungent sensation when eaten.Through construction of a pepper variome map,we examined the main groups that emerged during domestication and breeding of C.annuum,their relationships and temporal succession,and the molecular events underlying the main transitions.The results showed that the initial differentiation in fruit shape and pungency,increase in fruit weight,and transition from erect to pendent fruits,as well as the recent appearance of large,blocky,sweet fruits(bell peppers),were accompanied by strong selection/fixation of key alleles and introgressions in two large genomic regions.Furthermore,we identified Up,which encodes a BIG GRAIN protein involved in auxin transport,as a key domestication gene that controls erect vs pendent fruit orientation.The up mutation gained increased expression especially in the fruit pedicel through a 579-bp sequence deletion in its 5′upstream region,resulting in the phenotype of pendent fruit.The function of Up was confirmed by virus-induced gene silencing.Taken together,these findings constitute a cornerstone for understanding the domestication and differentiation of a key horticultural crop.展开更多
基金Supported by College Student Innovation Fund Project of Jilin University(2015821243)~~
文摘[Objective] This study was conducted to investigate cloning and expression of Pun1 gene controlling pungency of pepper. [Method] With Capsicum annuum L as a material, the cDNA sequence of Capunl gene was obtained, with a total length of 1 457 bp, coding 440 amino acids. [Result] Phylogenetic analysis showed that Capunl was closest to Pun1 of C. chinense, with a genetic distance of 0.019 3. Plant expression vector pCAM-Punl-GFP was constructed and transformed into to- bacco, and it was found that the protein coded by fusion gene Punl：：GFP was lo- cated on cell membrane. Prokaryotic expression vectors were constructed, and by SDS-PAGE and Western Blot detection, an induced protein with a molecular weight of 63 ku was obtained. It was found by real-time fluorescence quantitative expres- sion that Pun1 gene was expressed at the highest level 30 d after flowering, de- creased then, and could not be detected substantially 40 and 45 d after flowering. [Conclusion] This study provides information and reference for molecular regulation mechanism of Pun1 gene.
文摘An attempt was made in an effort to synthesize a series of capsaicinoids, most of which are synthesized in our laboratory and characterized to be completely new members of capsaicinoids. The Structure-pungency dependence are presented and discussed.
文摘The participation of O-methyltransferase (COMT) in phenylpropanoid-mediated capsaicinoid biosynthesis has long been proposed. Ferulic acid, a phenylpropanoid intermediate, is a precursor of capsaicinoid biosynthesis and is produced from caffeic acid by the action of COMT. As previously reported that silencing Comt expression caused a drastic decrease in capsaicinoid accumulation, it was presumed that a Comt loss-of-function mutation would cause loss of pungency in Capsicum. This hypothesis was tested by cloning Comt1 and Comt2 from the placenta tissue of the pungent cultivar Habanero. The phylogenetic analysis and comparison of critical amino-acid residues for enzyme function showed that the two COMTs had high similarity with the COMTs of other plant species. Moreover, as the two Comts were both expressed in placenta tissue and expressed prior to the accumulation of capsaicinoids, the two genes could be candidates for capsaicinoid biosynthesis. Second, Comt1 loss-of-function mutants were screened from the germplasm. A truncated Comt1 transcript was expressed in non-pungent pepper No.3341 caused by deletion of the genomic region. The predicted No.3341 COMT1 lacked His-265, which was absolutely necessary for enzymatic activity. Contrary to our expectations, the Comt1 mutation was not related to non-pungency of No.3341, as the deletion of Comt1 did not co-segregate with non-pungency in the F2 population obtained from crossing No.3341 with Habanero. This result was confirmed by screening several pungent accessions harboring the same Comt1 deletion mutation. Although the participation of COMT in phenylpropanoid-mediated capsaicinoid biosynthesis has long been proposed, our present study shows that Comt1 can not be a target for controlling fruit pungency.
基金supported by the National HighTechnology Research and Development Program of China (863 Program 2010AA065104)
文摘Ten capsaicin analogues were synthesized and their pungency degrees were determined through Scoville Organoleptic Test. The relationship between the structure and pungency degree of these capsaicin analogues was discussed. Then four of these capsaicin analogues with higher pungency degree were picked out and added to anti-biofouling paints as repellents to study their anti-biofouling performance by shallow sea buoyant raft hung-plate experimentation. The results showed that capsaicin and dihydrocapsaicin exhibited equally good anti-biofouling performance while nordihydrocapsaicin and N-vanillylnonanamide had poor anti-biofouling performance. Experimental results also showed that the paints with only 0.1% capsaicin or dihydrocapsaicin as repellent without any other biocides had also exhibited good anti-biofouling performance, which provided a new idea for developing novel, more environment-friendly and Cu20-free antifouling paints.
基金funded by the National Natural Science Foundation of China(Grant Nos.32202502,U21A20230,32070331,32102380 and 32072580)National Key Research and Development Program(Grant No.2018YFD1000800)+3 种基金the Key-Area Research and Development Program of Guangdong Province(Grant No.2022B0202080001)the Special Fund for Seed Industry of Guangdong Province Rural Revitalization Strategy(Grant No.2022-NPY00-024)Tibet Autonomous Region of Lhasa City Science and Technology Project(Grant No.LSKJ202310)the Science and Technology Project of Bijie City(Grant No.BKK2022-3)。
文摘The transcriptional cascade and regulatory loop play crucial roles in regulating plant-specialized metabolite biosynthesis.Capsaicinoids are unique to the genus Capsicum and confer a pungent flavor to its fruits.However,the transcriptional regulation of capsaicinoid biosynthesis remains largely unknown.In this study,two AP2/ERF transcription factors(TFs),CaERF102 and CaERF111,were characterized for their role in the capsaicinoid biosynthesis process.Expression analysis of two ERFs and capsaicinoid biosynthetic genes(CBGs)suggested that they were associated with capsaicinoid biosynthesis.Both ERFs encode nuclear-localized proteins and function as transcriptional activators through their C-terminal activation motifs.The two ERF TFs participated in capsaicinoid biosynthesis by directly activating the promoters of key CBGs,and this activation was significantly enhanced when CaMYC2 was co-expressed.Moreover,CaERF102 and CaERF111 were found to interact with CaMYC2.This study helps elucidate the AP2/ERF TF regulatory network that governs capsaicinoid biosynthesis in Capsicum species.
基金China National Key Technology Research and Development Program(2016YFD0100200,2016YFD0101700,2018YFD1000800,and 2020YFD1001100)National Natural Science Foundation of China(NSFC grants 31722048,31972411,3170110879,32102382,and 32102386)+5 种基金Central Public-interest Scientific Institution Basal Research Fund(Y2022GH04,IVF-BRF2021015)Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(CAAS-ASTIP-IVFCAAS)Earmarked Fund for the China Agriculture Research System(CARS-25)CAAS-GAAS Coordinated Innovation Project of the Chinese Academy of Agricultural Sciences(2019)Key Laboratory of Biology and Genetic Improvement of Horticultural Crops,Ministry of Agriculture and Rural Affairs,P.R.ChinaHorizon 2020 G2P-SOL project(grant no.677379)from the EU.
文摘Pepper(Capsicum spp.)is an important vegetable crop that provides a unique pungent sensation when eaten.Through construction of a pepper variome map,we examined the main groups that emerged during domestication and breeding of C.annuum,their relationships and temporal succession,and the molecular events underlying the main transitions.The results showed that the initial differentiation in fruit shape and pungency,increase in fruit weight,and transition from erect to pendent fruits,as well as the recent appearance of large,blocky,sweet fruits(bell peppers),were accompanied by strong selection/fixation of key alleles and introgressions in two large genomic regions.Furthermore,we identified Up,which encodes a BIG GRAIN protein involved in auxin transport,as a key domestication gene that controls erect vs pendent fruit orientation.The up mutation gained increased expression especially in the fruit pedicel through a 579-bp sequence deletion in its 5′upstream region,resulting in the phenotype of pendent fruit.The function of Up was confirmed by virus-induced gene silencing.Taken together,these findings constitute a cornerstone for understanding the domestication and differentiation of a key horticultural crop.
