Anthocyanin is one of water-soluble natural pigments widely existing in flowers, fruits, stems, leaves and seeds of plants, and it is the major factor conferring pink or red to the petals of Rosa rugose. MYB TFs play ...Anthocyanin is one of water-soluble natural pigments widely existing in flowers, fruits, stems, leaves and seeds of plants, and it is the major factor conferring pink or red to the petals of Rosa rugose. MYB TFs play an important role in the anthocyanin synthesis in plants. This work aimed to clone the MYB gene related to anthocyanin synthesis in the petals of Rosa rugose, and explore the relationship between them to lay a good foundation for gene engineering improvement of R. rugose. Based on the transcriptional data, a full-length cDNA sequence of MYB Gene, RrMYB113 (GenBank accession Nos MG720012), was cloned at the first time from the petals of Rosa rugose “Zi zhi” with RT-PCR and RACE methods. The full-length cDNA is 885 bp with an open reading frame of 654 bp, encoding 216 amino acids. The derived RrMYB113 protein has a molecular weight of 25,297.64 Da, a calculated pI of 9.61, a R2R3-MYB domain and bHLH binding domain, and it also has the signature motifs ((A/S/G)NDV and KPRPR(T/S)), thus belonging to Sg6 R2R3-MYB subfamily. In the secondary structure of RrMYB113 protein, there is 37.04% α-helix, 39.81% random coil, 14.81% extended peptide chain, and 8.33% β-corner. There is no transmembrane domain and no signal peptide cleavage site, seventeen Ser phosphorylation sites, fifteen Thr phosphorylation sites, four Tyr phosphorylation sites, and no O-glycosylation sites. The expression of RrMYB113 increased with the color deepening in petals, and it expressed at a higher level in petals than in other tissues of R. rugose “Zi zhi”. These results are meaningful to reveal that RrMYB113 might be an important regulator in anthocyanin biosynthesis and coloration in the petals of R. rugose.展开更多
In order to find a method to break dormancy of Rosa rugose seeds and interspecific hybrid seeds between Rosa rugose and Rosa hybrid quickly, and accelerate the breeding process of interspecific hybridization between R...In order to find a method to break dormancy of Rosa rugose seeds and interspecific hybrid seeds between Rosa rugose and Rosa hybrid quickly, and accelerate the breeding process of interspecific hybridization between Rosa rugose and Rosa hybrid, the influence of concentrated acid, seed maturity, GA3 (gibberellin) and low temperature (4°C) on seed germination of Rosa rugose from Muping was researched under aseptic condition. The results showed that aseptic germination can significantly shorten the germination time of Rosa rugose seeds and raise its germination ratio. Before inoculation, concentrated acid treatment greatly increased the germination rate and reduced the contamination rate of the seeds. The higher the degree of maturity of seeds is, the lower the germination rate would be, and the best time for seed to aseptic germination is 60 d after pollination. The addition of GA3 in 1/2MS medium could promote seeds germination better, and when the concentration of GA3 was 0.15 mg/L, the seed germination ratio was the highest;the germination time decreased and the seed germination ratio increased gradually as the treatment time at 4°C lasted longer.展开更多
As an important signal transduction protein, G protein beta subunit gene encoded by oligonucleotides plays an important role in many physiological, biochemical and environmental stresses in plants. In order to underst...As an important signal transduction protein, G protein beta subunit gene encoded by oligonucleotides plays an important role in many physiological, biochemical and environmental stresses in plants. In order to understand the action mode of G protein beta subunit gene, this paper cloned a Wd40 gene related to G protein beta subunit gene, named RrG-beta1, based on the R. rugose—transcriptome data, using Rosa rugose “Zi zhi” as experimental materials. The full length of cDNA of the gene was obtained by RT-PCR and RACE methods. The total length of this gene is 981 bp, and it encodes 326 amino acids. After bioinformatics analysis, the molecular formula C1601H2520N450O486S11 was predicted;the relative molecular weight was 36,201.00 Da;the theoretical isoelectric point PI value was 6.71;and its instability index was 30.44. The total average hydrophobic index was -0.847. In the secondary structure of RrG-beta1 protein, there are 17 α-helix, 131 Random coil, and 141 extended peptide chain. Gene Bank Blast results showed that the amino acid sequence encoded by RrG-beta1 was more than 90% homologous with the beta-like protein of Rosa chinensis, Fragaria, Malus, Pyrus, Prunus, Arabidopsis and tobacco, so it can be inferred that the RrG-beta1 Gene is guanine nucleotide-binding protein subunit beta-like protein. Fluorescence quantitative expression analysis of RrG-beta1 protein decreased with the development of flower color, and it was speculated that it could exert negative regulation effect on flower color. The leaf expression was highest in the tissue part, so it was inferred that the signal was transmitted through the stoma on the leaf.展开更多
In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was ...In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was constructed and the vector was introduced into tobacco with the agrobacterium-mediated method. PCR results showed that the RrGlu gene was integrated into the tobacco genome.展开更多
Plants have evolved and perfected a series of light receptors to feel the light at different bands and regulate the expression, modification and interaction of related genes in plants through signal transduction. So f...Plants have evolved and perfected a series of light receptors to feel the light at different bands and regulate the expression, modification and interaction of related genes in plants through signal transduction. So far, many photoreceptors have been identified in plants, UVR8 has recently been identified as a receptor for UV-B light. This paper cloned a WD40 gene related to UVR8 protein subunit, named RrRUP2, based on the Rosa rugose transcriptome data, using Rosa rugose “Zi zhi” as experimental materials. The full length of cDNA of the gene was obtained by RT-PCR and RACE methods. The total length of this gene is 1173 bp, and it encodes 390 amino acids. After bioinformatics analysis, the molecular formula C3415H5659N1173O1434S313 was predicted;the relative molecular weight was 96129.27 Da;the theoretical isoelectric point PI value was 5.00;and its instability index was 47.06. The total average hydrophobic index was 0.750. In the secondary structure of RrRUP2 protein, there are 10 α-helix, 45 β-helix, 181 Random coil, and 154 Extended strand. Gene Bank Blast results showed that the amino acid sequence encoded by RrRUP2 was more than 90% homologous with the RUP2 protein of Rosa chinensis, Fragaria, Malus, Pyrus, Prunus, Juglans, Arabidopsis and Tobacco, so it can be inferred that the RrRUP2 gene is a WD repeat-containing protein. Regarding to fluorescence quantitative expression analysis of RrRUP2, we find its experssion pattern is corresponded with the accumulation of anthocyanins.展开更多
文摘Anthocyanin is one of water-soluble natural pigments widely existing in flowers, fruits, stems, leaves and seeds of plants, and it is the major factor conferring pink or red to the petals of Rosa rugose. MYB TFs play an important role in the anthocyanin synthesis in plants. This work aimed to clone the MYB gene related to anthocyanin synthesis in the petals of Rosa rugose, and explore the relationship between them to lay a good foundation for gene engineering improvement of R. rugose. Based on the transcriptional data, a full-length cDNA sequence of MYB Gene, RrMYB113 (GenBank accession Nos MG720012), was cloned at the first time from the petals of Rosa rugose “Zi zhi” with RT-PCR and RACE methods. The full-length cDNA is 885 bp with an open reading frame of 654 bp, encoding 216 amino acids. The derived RrMYB113 protein has a molecular weight of 25,297.64 Da, a calculated pI of 9.61, a R2R3-MYB domain and bHLH binding domain, and it also has the signature motifs ((A/S/G)NDV and KPRPR(T/S)), thus belonging to Sg6 R2R3-MYB subfamily. In the secondary structure of RrMYB113 protein, there is 37.04% α-helix, 39.81% random coil, 14.81% extended peptide chain, and 8.33% β-corner. There is no transmembrane domain and no signal peptide cleavage site, seventeen Ser phosphorylation sites, fifteen Thr phosphorylation sites, four Tyr phosphorylation sites, and no O-glycosylation sites. The expression of RrMYB113 increased with the color deepening in petals, and it expressed at a higher level in petals than in other tissues of R. rugose “Zi zhi”. These results are meaningful to reveal that RrMYB113 might be an important regulator in anthocyanin biosynthesis and coloration in the petals of R. rugose.
文摘In order to find a method to break dormancy of Rosa rugose seeds and interspecific hybrid seeds between Rosa rugose and Rosa hybrid quickly, and accelerate the breeding process of interspecific hybridization between Rosa rugose and Rosa hybrid, the influence of concentrated acid, seed maturity, GA3 (gibberellin) and low temperature (4°C) on seed germination of Rosa rugose from Muping was researched under aseptic condition. The results showed that aseptic germination can significantly shorten the germination time of Rosa rugose seeds and raise its germination ratio. Before inoculation, concentrated acid treatment greatly increased the germination rate and reduced the contamination rate of the seeds. The higher the degree of maturity of seeds is, the lower the germination rate would be, and the best time for seed to aseptic germination is 60 d after pollination. The addition of GA3 in 1/2MS medium could promote seeds germination better, and when the concentration of GA3 was 0.15 mg/L, the seed germination ratio was the highest;the germination time decreased and the seed germination ratio increased gradually as the treatment time at 4°C lasted longer.
文摘As an important signal transduction protein, G protein beta subunit gene encoded by oligonucleotides plays an important role in many physiological, biochemical and environmental stresses in plants. In order to understand the action mode of G protein beta subunit gene, this paper cloned a Wd40 gene related to G protein beta subunit gene, named RrG-beta1, based on the R. rugose—transcriptome data, using Rosa rugose “Zi zhi” as experimental materials. The full length of cDNA of the gene was obtained by RT-PCR and RACE methods. The total length of this gene is 981 bp, and it encodes 326 amino acids. After bioinformatics analysis, the molecular formula C1601H2520N450O486S11 was predicted;the relative molecular weight was 36,201.00 Da;the theoretical isoelectric point PI value was 6.71;and its instability index was 30.44. The total average hydrophobic index was -0.847. In the secondary structure of RrG-beta1 protein, there are 17 α-helix, 131 Random coil, and 141 extended peptide chain. Gene Bank Blast results showed that the amino acid sequence encoded by RrG-beta1 was more than 90% homologous with the beta-like protein of Rosa chinensis, Fragaria, Malus, Pyrus, Prunus, Arabidopsis and tobacco, so it can be inferred that the RrG-beta1 Gene is guanine nucleotide-binding protein subunit beta-like protein. Fluorescence quantitative expression analysis of RrG-beta1 protein decreased with the development of flower color, and it was speculated that it could exert negative regulation effect on flower color. The leaf expression was highest in the tissue part, so it was inferred that the signal was transmitted through the stoma on the leaf.
文摘In order to lay a foundation for researching the function of Rosa rugose (R. rugosa) RrGlu gene, the RrGlu gene was amplified from the styles of R. rugosa “Tanghong”, a gene expression vector named PBI121-RrGlu was constructed and the vector was introduced into tobacco with the agrobacterium-mediated method. PCR results showed that the RrGlu gene was integrated into the tobacco genome.
文摘Plants have evolved and perfected a series of light receptors to feel the light at different bands and regulate the expression, modification and interaction of related genes in plants through signal transduction. So far, many photoreceptors have been identified in plants, UVR8 has recently been identified as a receptor for UV-B light. This paper cloned a WD40 gene related to UVR8 protein subunit, named RrRUP2, based on the Rosa rugose transcriptome data, using Rosa rugose “Zi zhi” as experimental materials. The full length of cDNA of the gene was obtained by RT-PCR and RACE methods. The total length of this gene is 1173 bp, and it encodes 390 amino acids. After bioinformatics analysis, the molecular formula C3415H5659N1173O1434S313 was predicted;the relative molecular weight was 96129.27 Da;the theoretical isoelectric point PI value was 5.00;and its instability index was 47.06. The total average hydrophobic index was 0.750. In the secondary structure of RrRUP2 protein, there are 10 α-helix, 45 β-helix, 181 Random coil, and 154 Extended strand. Gene Bank Blast results showed that the amino acid sequence encoded by RrRUP2 was more than 90% homologous with the RUP2 protein of Rosa chinensis, Fragaria, Malus, Pyrus, Prunus, Juglans, Arabidopsis and Tobacco, so it can be inferred that the RrRUP2 gene is a WD repeat-containing protein. Regarding to fluorescence quantitative expression analysis of RrRUP2, we find its experssion pattern is corresponded with the accumulation of anthocyanins.
