At present, the research about flower color of Rosa rugosa is a very inno-vative and practical study. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in ...At present, the research about flower color of Rosa rugosa is a very inno-vative and practical study. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1422bp, encoding 473 amino acids, designated as RrGT2, were isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT2 gene is C2334H3628N602O711S18, the relative molecular mass is 52,075.17 Da, and the theoretical isoelectric point is pI = 4.76. The result of the RrGT2 protein 3D model construction showed that it had the highest homology with the UDP-glycosyltransferase 74F2 protein model in the database (39.53%). Sequence alignments with the NCBI database showed that the RrGT2 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT2 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT2 enzyme activity. RrGT2 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of antho-cyanins. Therefore, we speculated that glycosylation of RrGT2 plays a crucial role in anthocyanin biosynthesis in R. rugosa.展开更多
Virus-induced gene silencing (VIGS) technique, which is developed in recent years, is a rapid identification of plant gene function from reverse genetics. It is a manifestation of post-transcriptional gene silencing m...Virus-induced gene silencing (VIGS) technique, which is developed in recent years, is a rapid identification of plant gene function from reverse genetics. It is a manifestation of post-transcriptional gene silencing mechanism. Compared with the traditional transgenic technology, VIGS is a transient expression system, which can achieve good results in a short time. At present, it is widely used to study the function of plant genes, but most of them are model plants, and the experiments are carried out always in the indoor environment with controlled light and temperature conditions. In this study, we creatively provided a method to establish VIGS system using perennial Rosa plants as experimental materials under field conditions. The recombinant virus vector was constructed with RrGT1 gene as reporter gene and modified TRV-GFP virus as vector, and the perennial R. rugosa “Zizhi” and R. davurica were used as experimental verification materials. According to the growth conditions of Rosa plants, the natural environment in the field and the optimal conditions for the occurrence of VIGS, the technical problems such as the confirmation of the inoculation period, the preparation of the infective fluid, the inoculation technology of the virus vector and the light and temperature conditions of plant materials cultured after inoculation were solved one by one. When the RrGT1 gene was silenced, the Rosa plants showed a pale petal color phenotype. By detection, it was found that the expression of endogenous RrGT1 gene was significantly down-regulated, and the content of all anthocyanins also decreased significantly. Therefore, we believed that the attempt to establish VIGS system in perennial Rosa plants under field conditions was very successful.展开更多
The species and contents of anthocyanins in plant petals can make plants appear pink, red, violet and blue, etc., and play a major role in the coloration of plants. In this study, the species and contents of anthocyan...The species and contents of anthocyanins in plant petals can make plants appear pink, red, violet and blue, etc., and play a major role in the coloration of plants. In this study, the species and contents of anthocyanins in the petals of three R. rugosa hybrid cultivars, R. rugosa “Zizhi”, R. rugosa “Fenzizhi” and R. rugosa “Baizizhi”, were analyzed, and the direct cause of the differences in flower color of three R. rugosa hybrid cultivars was inferred. This paper provides a reference for the coloration mechanism and flower color breeding of R. rugosa. The specific methods are as follows: the petals of five flowering stages of three R. rugosa hybrid cultivars were used as materials, and the types and contents of anthocyanins contained in them were qualitatively and quantitatively analyzed by high performance liquid chromatography (HPLC). The same six kinds of anthocyanins were identified in R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, mainly based on the diglycoside of paeoniflorin and cyanidin. The relative contents of the two anthocyanins were higher at budding stage and initial opening stage. In the different flowering stages of R. rugosa “Zizhi”, the content of Pn3G5G was up to 4280.84 ± 20.82 μg·g-1, and the content of Cy3G5G was up to 789.41 ± 1.21 μg·g-1. In R. rugosa “Fenzizhi”, the highest content of Pn3G5G reached 1293.50 ± 17.64 μg·g-1, and the content of Cy3G5G was up to 358.86 ± 3.94 μg·g-1. It could be speculated that the difference in the contents of Pn3G5G and Cy3G5G was the main reason for the difference in coloration between the petals of R. rugosa “Zizhi” and R. rugosa “Fenzizhi”. A total of five species of anthocyanins were identified in R. rugosa “Baizizhi” and their contents were relatively low. Compared with R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, the presence of Cy3G was not detected. Therefore, we speculated that the two reasons above might be responsible for the visual white flowers of R. rugosa “Baizizhi”.展开更多
Based on the transcriptome of Rosa rugosa, one anthocyanin-promoting R2R3-MYB gene, RrMYB10.1 (Accession Nos:MH717244), was cloned from the petals of Rosa rugosa ‘Zizhi’. Sequence analysis results showed that RrMYB1...Based on the transcriptome of Rosa rugosa, one anthocyanin-promoting R2R3-MYB gene, RrMYB10.1 (Accession Nos:MH717244), was cloned from the petals of Rosa rugosa ‘Zizhi’. Sequence analysis results showed that RrMYB10.1 had a full length opening reading frame of 747bp, encoding 249 amino acids. Sequence analysis revealed that RrMYB10.1 contained the conserved R2R3-MYB domain, two atypical anthocyanin-promoting motifs and a conserved amino acid signature for the interaction with bHLH protein. The results of phylogenic tree revealed that RrMYB10.1 showed high homology with other anthocyanin-promoting proteins in Rosacea, and sharing the highest identity (98.39%) with RhMYB10. RT-PCR results showed that RrMYB10.1 was mainly expressed in petals among various tissues and expressed significantly higher in petals in bud stage than in opening period. To sum up, these results showed that RrMYN10.1 may play a key role in regulating anthocyanin concentration, thus providing a certain foundation on regulating flower color formation in Rosa rugosa.展开更多
Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDN...Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1161 bp, encoding 386 amino acids, designated as RrGT1, was isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT1 gene is C1879H2964N494O556S14, the relative molecular mass is 41,820.02 Da, and the theoretical isoelectric point is pI = 5.03. The result of the RrGT1 protein 3D model construction showed that it had the highest homology with the UDP-glucose: anthocyanidin 3-O-glucosyltransferase protein model in the database (47.01%). Sequence alignments with the NCBI database showed that the RrGT1 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT1 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT1 enzyme activity. RrGT1 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of anthocyanins. Therefore, we speculated that glycosylation of RrGT1 plays a crucial role in anthocyanin biosynthesis in R. rugosa.展开更多
文摘At present, the research about flower color of Rosa rugosa is a very inno-vative and practical study. Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1422bp, encoding 473 amino acids, designated as RrGT2, were isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT2 gene is C2334H3628N602O711S18, the relative molecular mass is 52,075.17 Da, and the theoretical isoelectric point is pI = 4.76. The result of the RrGT2 protein 3D model construction showed that it had the highest homology with the UDP-glycosyltransferase 74F2 protein model in the database (39.53%). Sequence alignments with the NCBI database showed that the RrGT2 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT2 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT2 enzyme activity. RrGT2 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of antho-cyanins. Therefore, we speculated that glycosylation of RrGT2 plays a crucial role in anthocyanin biosynthesis in R. rugosa.
文摘Virus-induced gene silencing (VIGS) technique, which is developed in recent years, is a rapid identification of plant gene function from reverse genetics. It is a manifestation of post-transcriptional gene silencing mechanism. Compared with the traditional transgenic technology, VIGS is a transient expression system, which can achieve good results in a short time. At present, it is widely used to study the function of plant genes, but most of them are model plants, and the experiments are carried out always in the indoor environment with controlled light and temperature conditions. In this study, we creatively provided a method to establish VIGS system using perennial Rosa plants as experimental materials under field conditions. The recombinant virus vector was constructed with RrGT1 gene as reporter gene and modified TRV-GFP virus as vector, and the perennial R. rugosa “Zizhi” and R. davurica were used as experimental verification materials. According to the growth conditions of Rosa plants, the natural environment in the field and the optimal conditions for the occurrence of VIGS, the technical problems such as the confirmation of the inoculation period, the preparation of the infective fluid, the inoculation technology of the virus vector and the light and temperature conditions of plant materials cultured after inoculation were solved one by one. When the RrGT1 gene was silenced, the Rosa plants showed a pale petal color phenotype. By detection, it was found that the expression of endogenous RrGT1 gene was significantly down-regulated, and the content of all anthocyanins also decreased significantly. Therefore, we believed that the attempt to establish VIGS system in perennial Rosa plants under field conditions was very successful.
文摘The species and contents of anthocyanins in plant petals can make plants appear pink, red, violet and blue, etc., and play a major role in the coloration of plants. In this study, the species and contents of anthocyanins in the petals of three R. rugosa hybrid cultivars, R. rugosa “Zizhi”, R. rugosa “Fenzizhi” and R. rugosa “Baizizhi”, were analyzed, and the direct cause of the differences in flower color of three R. rugosa hybrid cultivars was inferred. This paper provides a reference for the coloration mechanism and flower color breeding of R. rugosa. The specific methods are as follows: the petals of five flowering stages of three R. rugosa hybrid cultivars were used as materials, and the types and contents of anthocyanins contained in them were qualitatively and quantitatively analyzed by high performance liquid chromatography (HPLC). The same six kinds of anthocyanins were identified in R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, mainly based on the diglycoside of paeoniflorin and cyanidin. The relative contents of the two anthocyanins were higher at budding stage and initial opening stage. In the different flowering stages of R. rugosa “Zizhi”, the content of Pn3G5G was up to 4280.84 ± 20.82 μg·g-1, and the content of Cy3G5G was up to 789.41 ± 1.21 μg·g-1. In R. rugosa “Fenzizhi”, the highest content of Pn3G5G reached 1293.50 ± 17.64 μg·g-1, and the content of Cy3G5G was up to 358.86 ± 3.94 μg·g-1. It could be speculated that the difference in the contents of Pn3G5G and Cy3G5G was the main reason for the difference in coloration between the petals of R. rugosa “Zizhi” and R. rugosa “Fenzizhi”. A total of five species of anthocyanins were identified in R. rugosa “Baizizhi” and their contents were relatively low. Compared with R. rugosa “Zizhi” and R. rugosa “Fenzizhi”, the presence of Cy3G was not detected. Therefore, we speculated that the two reasons above might be responsible for the visual white flowers of R. rugosa “Baizizhi”.
文摘Based on the transcriptome of Rosa rugosa, one anthocyanin-promoting R2R3-MYB gene, RrMYB10.1 (Accession Nos:MH717244), was cloned from the petals of Rosa rugosa ‘Zizhi’. Sequence analysis results showed that RrMYB10.1 had a full length opening reading frame of 747bp, encoding 249 amino acids. Sequence analysis revealed that RrMYB10.1 contained the conserved R2R3-MYB domain, two atypical anthocyanin-promoting motifs and a conserved amino acid signature for the interaction with bHLH protein. The results of phylogenic tree revealed that RrMYB10.1 showed high homology with other anthocyanin-promoting proteins in Rosacea, and sharing the highest identity (98.39%) with RhMYB10. RT-PCR results showed that RrMYB10.1 was mainly expressed in petals among various tissues and expressed significantly higher in petals in bud stage than in opening period. To sum up, these results showed that RrMYN10.1 may play a key role in regulating anthocyanin concentration, thus providing a certain foundation on regulating flower color formation in Rosa rugosa.
文摘Glycosylation modification fulfills an important role in increasing the stability and solubility of anthocyanin in plants. In this study, based on the transcriptional database of R. rugosa, a gene with full length cDNA of 1161 bp, encoding 386 amino acids, designated as RrGT1, was isolated from flowers of R. rugosa ‘Zizhi’ and then functionally characterized. According to online software prediction, the molecular formula of the protein encoded by the RrGT1 gene is C1879H2964N494O556S14, the relative molecular mass is 41,820.02 Da, and the theoretical isoelectric point is pI = 5.03. The result of the RrGT1 protein 3D model construction showed that it had the highest homology with the UDP-glucose: anthocyanidin 3-O-glucosyltransferase protein model in the database (47.01%). Sequence alignments with the NCBI database showed that the RrGT1 protein is a member of the GTB superfamily. Homology analysis revealed that the coding regions of RrGT1 was highly specific among different species, but still had typical conserved amino acid residues called PSPG that are crucial for RrGT1 enzyme activity. RrGT1 transcripts were detected in five flowering stages and seven tissues of R. rugosa ‘Zizhi’, R. rugosa ‘Fenzizhi’ and R. rugosa ‘Baizizhi’, and their expression patterns corresponded with the accumulation of anthocyanins. Therefore, we speculated that glycosylation of RrGT1 plays a crucial role in anthocyanin biosynthesis in R. rugosa.
