Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response...Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response in Arabidopsis thaliana.However,the issue of the induction of the VIN3-LIKE genes in wintersweet(Chimonanthus praecox)has been largely neglected.In the present study,we explored the molecular regulation of the PHD type finger protein-encoding gene CpVIL2 in relation to the growth and development of wintersweet in Arabidopsis.In wintersweet,quantitative real-time PCR(qRT-PCR)analysis showed that the relative expression of CpVIL2-As2i(intron-retained alternatively spliced in the second intron)was extremely higher in the pistils than in the other tissues.And the relative CpVIL2-As2i expression in flower buds(FBs)treated at 8°C was higher than that of FBs in December,2016 under natural conditions,which was not detected in non-flowering FBs at 16°C.In Arabidopsis,the expression patterns of the CpVIL2-As2i gene were detected at first in CpVIL2-As2i pro::GUS(β-glucuronidase)lines,with predominantly higher expression in flowers and inflorescence.Meanwhile,the hormone-induced expression profiles of the CpVIL2-As2i promoter were confirmed using exogenous induction by abscisic acid(ABA)and indole acetic acid(IAA)phytohormones,where the GUS enzyme activity obviously decreased compared with that of control.In comparison with Arabidopsis/Col-0,early flowering was detected in ectopic 35S::CpVIL2-As2i lines.Overall,these results demonstrated the function of the CpVIL2-As2i gene,at the same time,provided us with new insights into the molecular mechanisms of early flowering and complex regulatory networks of vernalization in wintersweet.展开更多
[Objective] The aim was to study the cDNA cloning of SMAT gene from Chimonanthus praecox and its expression. [Method] A novel cDNA was cloned by PT-PCR using the total RNA as template, and amplified to the desirable s...[Objective] The aim was to study the cDNA cloning of SMAT gene from Chimonanthus praecox and its expression. [Method] A novel cDNA was cloned by PT-PCR using the total RNA as template, and amplified to the desirable size. The RT-PCR products were reclaimed and transformed into E. coli DH 5o together with the PMD18-T vector after ligating by T-A cloning. Identified by colony PCR and EcoRI and Notl digestion, the recombinant plasmid with target gene was screened out and conducted the sequence analysis. [Result] Results of the sequence analysis showed that the ORF fragment of the SAMT cDNA was successfully cloned from Chimonanthus praecox gene, with the length of 1 196 bp and encoding 380 amino acids fragment which shared 99.2% homology to that of previously reported SAMT cDNA from Chimonanthus praecox （ABU88887）. The SAMT cDNA fragment was sub-cloned into the prokaryotic expression vector PGEX1-4T-1, and the obtained re- combinant plasmid was named PGSAMT. After inducting by 0.01mol/L IPTG, the re- sult of the SDS-PAGE analysis showed that the molecular weight of the fusion ex- pression SAMT protein was about 66 kDa, which was close to the predicted fusion protein derived from the 26 kDa GST band and 42.3 kDa SAMT gene of Chimo- nanthus praecox encoded protein. [Conclusion] This study successfully cloned and expressed the SAMT gene of Chimonanthus praecox.展开更多
Chimonanthus praecox is an important ornamental plant and cut flower material in China.It blooms in the freezing winter and its flower emits charming fragrance.However,in different region the flowering time is variabl...Chimonanthus praecox is an important ornamental plant and cut flower material in China.It blooms in the freezing winter and its flower emits charming fragrance.However,in different region the flowering time is variable.In order to understand the flowering mechanism of Ch.praecox in the winter,we studied the flower bud differentiation in Spring City-Kunming using paraffin sectioning method in the present study.Meanwhile we compared the differentiation process difference from different regions.It was found that the temperature is the key factor for its flower bud differentiation and blossom of Ch.praecox.In the process of bud differentiation,the temperature 20℃was the optimum for inducing changes from vegetative axillary buds to reproductive buds and subsequent morphological differentiation in Ch.praecox.Furthermore in the first three differentiation periods—tepal primordial stage,staminal primordial stage and pistil primordial stage,Kunming took the shortest time to finish the process due to very rapid temperature rise to 20℃,whereas,in Zhengzhou the time for these differentiations was the longest,which may be caused by the slow temperature rise.After May,the high temperature stress forced the flower buds into the first long dormant period in all regions except Kunming.In Kunming,the average temperature was only 20–25℃,so the flower bud continued to differentiate.In all regions,Kunming is the first to complete whole flower bud differentiation even on the early August,and started the second dormancy very early but very long.In the other regions,the plants went through a shorter dormancy and the low temperature broke the dormancy rapidly.Contrarily the plants of Kunming spent a longer period for the low temperature.Thus,the low temperature less than 10℃is a key factor to breaking the second dormancy.Surely the regular effects of temperature on flower bud differentiation and blossom is very helpful for florescence regulation of Ch.praecox.展开更多
Objective] To analyze the aroma constituents of essential oils from Chi-monanthus praecox. [Method] Extracted by microwave-assisted hydrodistil ation （MAHD）, aroma constituents of essential oils of Chimonanthus prae...Objective] To analyze the aroma constituents of essential oils from Chi-monanthus praecox. [Method] Extracted by microwave-assisted hydrodistil ation （MAHD）, aroma constituents of essential oils of Chimonanthus praecox flowers were analyzed by gas chromatography-mass spectrometry （GC-MS）. Normalization method was used to determine the constituents quantitatively. [Result] Total y 27 compounds were identified, accounting for 98.85% of total amounts. The main aroma con-stituents are terpenes, aldehydes, esters, alcohols and hydrocarbons. Main compo-nents with relative percentage over 5% are germacrene D （25.62%）, bornyl acetate （16.71%）, caryophyl ene （10.51%）, cis-α-ocimene （5.18%）, γ-elemene （8.05%）, β-linalool （5.01%）, 1-ethenyl-1-methyl-2,4-bis （1-methylethenyl）-cyclohexane （7.18%） and 2,3,4,4α,5,6-hexahydro-1,4αdimethyl-7-（1-methylethyl）-naphthalene （5.20%）. [Conclu-sion] The study provided a theoretical reference for the development and utilization of Chimonanthus praecox resources.展开更多
利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在...利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在启动子的基本元件TATA-box和CAAT-box及多个与植物非生物胁迫相关的响应元件。在烟草叶片中的瞬时表达结果表明这两个启动子序列均具备驱动报告基因GUS表达的功能。展开更多
通过对影响AFLP反应体系主要因素的优化,建立了蜡梅的AFLP反应体系,并筛选出了适宜该体系分析的引物.结果表明,20μL蜡梅AFLP最佳酶切体系为模板600 ng DNA,3 U Pst I和3 U Mse I,在37℃下双酶切2 h;在20μL最佳连接体系中酶切产物为15...通过对影响AFLP反应体系主要因素的优化,建立了蜡梅的AFLP反应体系,并筛选出了适宜该体系分析的引物.结果表明,20μL蜡梅AFLP最佳酶切体系为模板600 ng DNA,3 U Pst I和3 U Mse I,在37℃下双酶切2 h;在20μL最佳连接体系中酶切产物为15μL,3 U T4连接酶,0.25μmol.L-1 Pst I接头,2.5μmol.L-1 Mse I接头,1μL 10×T4 Buffer,在22℃下连接10 h;在20μL最佳预扩反应体系中稀释10倍的连接产5μL,2.0mmol.L-1 Mg2+,2 U Taq酶,300μmol.L-1dNTP,0.5μmol.L-1 Pst I和Mse I引物(P+AGA/M+ATC);在20μL最佳选择性扩增反应体系中5μL稀释20倍的预扩增产物,2.0 mmol.L-1的Mg2+,2 U Taq酶,300μmol.L-1 dNTP,0.5μmol.L-1 Pst I和Mse I引物(P+AGA/M+ATC).最后,利用上面的体系筛选出了96对适宜于蜡梅AFLP分析的引物.展开更多
基金supported by the Grants from the Natural Science Foundation of Chongqing(No.cstc2020jcyj-msxmX1014)Fundamental Research Funds for the Central Universities(No.XDJK2020B059)National Natural Science Foundation of China(Grant No.31971711).
文摘Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response in Arabidopsis thaliana.However,the issue of the induction of the VIN3-LIKE genes in wintersweet(Chimonanthus praecox)has been largely neglected.In the present study,we explored the molecular regulation of the PHD type finger protein-encoding gene CpVIL2 in relation to the growth and development of wintersweet in Arabidopsis.In wintersweet,quantitative real-time PCR(qRT-PCR)analysis showed that the relative expression of CpVIL2-As2i(intron-retained alternatively spliced in the second intron)was extremely higher in the pistils than in the other tissues.And the relative CpVIL2-As2i expression in flower buds(FBs)treated at 8°C was higher than that of FBs in December,2016 under natural conditions,which was not detected in non-flowering FBs at 16°C.In Arabidopsis,the expression patterns of the CpVIL2-As2i gene were detected at first in CpVIL2-As2i pro::GUS(β-glucuronidase)lines,with predominantly higher expression in flowers and inflorescence.Meanwhile,the hormone-induced expression profiles of the CpVIL2-As2i promoter were confirmed using exogenous induction by abscisic acid(ABA)and indole acetic acid(IAA)phytohormones,where the GUS enzyme activity obviously decreased compared with that of control.In comparison with Arabidopsis/Col-0,early flowering was detected in ectopic 35S::CpVIL2-As2i lines.Overall,these results demonstrated the function of the CpVIL2-As2i gene,at the same time,provided us with new insights into the molecular mechanisms of early flowering and complex regulatory networks of vernalization in wintersweet.
文摘[Objective] The aim was to study the cDNA cloning of SMAT gene from Chimonanthus praecox and its expression. [Method] A novel cDNA was cloned by PT-PCR using the total RNA as template, and amplified to the desirable size. The RT-PCR products were reclaimed and transformed into E. coli DH 5o together with the PMD18-T vector after ligating by T-A cloning. Identified by colony PCR and EcoRI and Notl digestion, the recombinant plasmid with target gene was screened out and conducted the sequence analysis. [Result] Results of the sequence analysis showed that the ORF fragment of the SAMT cDNA was successfully cloned from Chimonanthus praecox gene, with the length of 1 196 bp and encoding 380 amino acids fragment which shared 99.2% homology to that of previously reported SAMT cDNA from Chimonanthus praecox （ABU88887）. The SAMT cDNA fragment was sub-cloned into the prokaryotic expression vector PGEX1-4T-1, and the obtained re- combinant plasmid was named PGSAMT. After inducting by 0.01mol/L IPTG, the re- sult of the SDS-PAGE analysis showed that the molecular weight of the fusion ex- pression SAMT protein was about 66 kDa, which was close to the predicted fusion protein derived from the 26 kDa GST band and 42.3 kDa SAMT gene of Chimo- nanthus praecox encoded protein. [Conclusion] This study successfully cloned and expressed the SAMT gene of Chimonanthus praecox.
基金funded by Talents Introduction Plan of Yunnan Province-"High-End Foreign Experts"Program(Grant No.000019)。
文摘Chimonanthus praecox is an important ornamental plant and cut flower material in China.It blooms in the freezing winter and its flower emits charming fragrance.However,in different region the flowering time is variable.In order to understand the flowering mechanism of Ch.praecox in the winter,we studied the flower bud differentiation in Spring City-Kunming using paraffin sectioning method in the present study.Meanwhile we compared the differentiation process difference from different regions.It was found that the temperature is the key factor for its flower bud differentiation and blossom of Ch.praecox.In the process of bud differentiation,the temperature 20℃was the optimum for inducing changes from vegetative axillary buds to reproductive buds and subsequent morphological differentiation in Ch.praecox.Furthermore in the first three differentiation periods—tepal primordial stage,staminal primordial stage and pistil primordial stage,Kunming took the shortest time to finish the process due to very rapid temperature rise to 20℃,whereas,in Zhengzhou the time for these differentiations was the longest,which may be caused by the slow temperature rise.After May,the high temperature stress forced the flower buds into the first long dormant period in all regions except Kunming.In Kunming,the average temperature was only 20–25℃,so the flower bud continued to differentiate.In all regions,Kunming is the first to complete whole flower bud differentiation even on the early August,and started the second dormancy very early but very long.In the other regions,the plants went through a shorter dormancy and the low temperature broke the dormancy rapidly.Contrarily the plants of Kunming spent a longer period for the low temperature.Thus,the low temperature less than 10℃is a key factor to breaking the second dormancy.Surely the regular effects of temperature on flower bud differentiation and blossom is very helpful for florescence regulation of Ch.praecox.
基金Supported by the Research Fund from Guizhou Academy of Agricultural Science(2013009)Innovation Capacity Platform Construction Project of Guizhou Science and Technology Department(20104008)~~
文摘Objective] To analyze the aroma constituents of essential oils from Chi-monanthus praecox. [Method] Extracted by microwave-assisted hydrodistil ation （MAHD）, aroma constituents of essential oils of Chimonanthus praecox flowers were analyzed by gas chromatography-mass spectrometry （GC-MS）. Normalization method was used to determine the constituents quantitatively. [Result] Total y 27 compounds were identified, accounting for 98.85% of total amounts. The main aroma con-stituents are terpenes, aldehydes, esters, alcohols and hydrocarbons. Main compo-nents with relative percentage over 5% are germacrene D （25.62%）, bornyl acetate （16.71%）, caryophyl ene （10.51%）, cis-α-ocimene （5.18%）, γ-elemene （8.05%）, β-linalool （5.01%）, 1-ethenyl-1-methyl-2,4-bis （1-methylethenyl）-cyclohexane （7.18%） and 2,3,4,4α,5,6-hexahydro-1,4αdimethyl-7-（1-methylethyl）-naphthalene （5.20%）. [Conclu-sion] The study provided a theoretical reference for the development and utilization of Chimonanthus praecox resources.
文摘利用hiTAIL-PCR法得到了2个蜡梅(Chimonanthus praecox)非特异性脂转移蛋白(non-specific lipid transfer protein)基因家族成员CpLTP3和CpLTP4翻译起始位点上游启动子序列,长度分别为1298bp和838bp。生物信息学分析表明2个序列均存在启动子的基本元件TATA-box和CAAT-box及多个与植物非生物胁迫相关的响应元件。在烟草叶片中的瞬时表达结果表明这两个启动子序列均具备驱动报告基因GUS表达的功能。
