Objective To clone the full-length cDNA of 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGR) from Aquilaria sinensis(AsHMGR1) and to analyze its expression profile in different tissues and in response to different...Objective To clone the full-length cDNA of 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGR) from Aquilaria sinensis(AsHMGR1) and to analyze its expression profile in different tissues and in response to different treatments.HMGR is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate pathway.Methods RT-PCR and RACE were used to clone the full-length cDNA of HMGR from A.sinensis based on the conserved HMGR gene fragments.The bioinformatic analysis was performed on its nucleic acid and protein sequence.The expression profile of AsHMGR1 in different tissues and in response to different treatments was analyzed by quantitative RT-PCR.Results The full-length AsHMGR1 cDNA was 2026 bp,containing a 1719 bp open reading frame which encoded a protein of 572 amino acids.Amino acid sequence homology alignment and phylogenetic analysis demonstrated that AsHMGR1 belonged to the HMGR gene family.The detection of tissue expression patterns showed that AsHMGR1 was mainly expressed in the stem,followed by roots and branches.AsHMGR1 could be stimulated by methyl jasmonate and H2O2to varying degrees in a time-dependent manner.Conclusion These data will provide a foundation for further investigation on AsHMGR1 functions and regulatory mechanisms in sesquiterpene synthesis in A.sinensis.展开更多
Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum ch...Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0%to 2.2%and 5.0%for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D).The highest regeneration rate(34.6%)was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine(BA).The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant(2n=12)with only one haploid plant(n=6).Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture of B.chinense.展开更多
基金National Natural Science Funds of China(3110022081173539)the National Science and Technology Support Project(2011BAI01B07)
文摘Objective To clone the full-length cDNA of 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGR) from Aquilaria sinensis(AsHMGR1) and to analyze its expression profile in different tissues and in response to different treatments.HMGR is the first rate-limiting enzyme for sesquiterpene synthesis in the mevalonate pathway.Methods RT-PCR and RACE were used to clone the full-length cDNA of HMGR from A.sinensis based on the conserved HMGR gene fragments.The bioinformatic analysis was performed on its nucleic acid and protein sequence.The expression profile of AsHMGR1 in different tissues and in response to different treatments was analyzed by quantitative RT-PCR.Results The full-length AsHMGR1 cDNA was 2026 bp,containing a 1719 bp open reading frame which encoded a protein of 572 amino acids.Amino acid sequence homology alignment and phylogenetic analysis demonstrated that AsHMGR1 belonged to the HMGR gene family.The detection of tissue expression patterns showed that AsHMGR1 was mainly expressed in the stem,followed by roots and branches.AsHMGR1 could be stimulated by methyl jasmonate and H2O2to varying degrees in a time-dependent manner.Conclusion These data will provide a foundation for further investigation on AsHMGR1 functions and regulatory mechanisms in sesquiterpene synthesis in A.sinensis.
基金National Key Project of Scientific and Technical Supporting Programs Funded by the Ministry of Science & Technology of China (2006BAI09B01)Research Fund for the Doctoral Program of Advance Education of China (20070023094)Beijing Natural Science Foundation (6082020)
文摘Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0%to 2.2%and 5.0%for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D).The highest regeneration rate(34.6%)was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine(BA).The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant(2n=12)with only one haploid plant(n=6).Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture of B.chinense.
