Rose(Rosa hybrida)is widely used for cut flowers and as garden plants.Stable and efficient transformation system is required for functional genomics of rose.Here,we established an efficient transformation method for r...Rose(Rosa hybrida)is widely used for cut flowers and as garden plants.Stable and efficient transformation system is required for functional genomics of rose.Here,we established an efficient transformation method for rose using Agrobacterium tumefaciens-mediated transformation of embryogenic callus.Expanding rose leaves were used as explants to induce somatic embryos,which were subjected to transformation with A.tumefaciens strain GV3101 using Green Fluorescence Protein(GFP)as a marker gene.It took about 8 months to generate transgenic shoots from embryogenic callus.PCR,RT-PCR,Southern and Western blotting,as well as stereoscopic fluorescence microscopy analysis demonstrated that GFP transgenes integrated stably into the rose genome.According to our data,a transformation efficiency of up to 6%can be achieved by following this optimized protocol.展开更多
Meiotic recombination and the resulting novel allele combinations are fundamental to plant breeding and the identification of superior hybrids.However,the rate of meiotic crossovers is naturally suppressed below its p...Meiotic recombination and the resulting novel allele combinations are fundamental to plant breeding and the identification of superior hybrids.However,the rate of meiotic crossovers is naturally suppressed below its potential ability,which prevents plant breeding efficiency.Nine suppressors of meiotic recombination have been identified in the model plant Arabidopsis and in other crop species.Mutations in these genes can lead to increased recombination frequency and could therefore potentially be used to create hyper-recombinant lines for ornamental breeding.In Gerbera hybrida,the anti-crossover factors remain elusive.In this study,we isolated and cloned TOP3αfrom flower buds of G.hybrida,and it encoded 935 amino acids with three conserved domains TOPRIM,TOP1Ac and zf-GR.Moreover,TOP3αwas the highest expressed at the flower bud stage,which coincided with the occurrence of meiotic recombination,suggesting that TOP3αis associated with the regulation of meiotic recombination in G.hybrida.展开更多
Gerbera Hybrida is one of the important cut flowers across the world.The novel traits are the primarily market requirements and the breeding targets,mainly determined by the degree of genetic variation after hybridiza...Gerbera Hybrida is one of the important cut flowers across the world.The novel traits are the primarily market requirements and the breeding targets,mainly determined by the degree of genetic variation after hybridization.However,meiotic recombination is highly conserved in most eukaryotes which suppressed the crossover formation and limited the genetic diversity.Recently,several meiotic recombination suppressors have been identified and characterized in plants,whereas it remains elusive in G.hybrida.In order to characterize the expression patterns of these suppressors in G.hybrida,20 candidate reference genes were identified from the transcriptome datasets of G.hybrida,and their expression stabilities during plant development were evaluated by geNorm,NormFinder and BestKeeper.Although the most stable reference genes were variable in different softwares,comprehensive ranking revealed that PGK2 was the most stable reference gene and GAPDH was the most unstable one.The expression patterns of FANCM,FIGL1,RECQ4,RM1,and FLIP further validated that PGK2 was suitable for normalization of gene expression.Our study identified a reliable reference gene for gene expression during meiotic recombination,and provided functional insights into meiotic recombination suppressors in G.hybrida.展开更多
To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron...To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing -glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600= 0.50.8 for 20 min and co-culture in darkness under 23 C on medium with 1/2 MS salts and 300 mol稬1 AS for 3 d.展开更多
In our previous study,a gene predicted to encode a Tyrosine aminotransferase( TAT) was found to be significantly up-regulated in root rot diseased Gerbera by transcriptome sequencing. To confirm the genes and investig...In our previous study,a gene predicted to encode a Tyrosine aminotransferase( TAT) was found to be significantly up-regulated in root rot diseased Gerbera by transcriptome sequencing. To confirm the genes and investigate the function,we cloned the gene by RT-PCR and then conduct bioinformatic analyses. In this study,a 1 537 bp long c DNA sequence of this gene( named as Gh TAT) was firstly cloned,which contained a coding region of 1 233 bp,which was predicted to encode a protein of 410 amino acids. Bioinformatic analysis showed that the Gh TAT was a stable hydrophobic protein without signal peptide. Subcellular location prediction result indicated that this protein located in chloroplast,which is the biosynthesis position of tyrosine and the derived products of tyrosine biosynthesis pathway. Moreover,typical Tyrosine aminotransferase domain was found in this protein,indicating that it is a TAT. According to the TAT-based phylogenetic analysis and similarity analysis,the closest relationship and highest similarity was found between Gh TAT and Halianthus annuus TAT,which again verified the TAT property of Gh TAT. Tyrosine aminotransferase( TAT) is the first enzyme in tyrosine biosynthesis pathway,whose products include many antioxidant substances such as tocopherols and tocotrienols. The up-regulation of Gh TAT in root rot diseased gerbera suggests that it may play an important role in response to the root rot pathogen infection. In addition,60 phosphorylation sites( accounting for 14. 6%) were found in this protein,suggesting that the expression of this protein and its encoding gene were greatly influenced by the phosphorylation reactions.展开更多
Effect of explant, site of leaflet, induction period in the dark and combinations of plant growth regulators on direct adventitious bud induction and plant regeneration of Rosa hybrida Samantha was investigated. The r...Effect of explant, site of leaflet, induction period in the dark and combinations of plant growth regulators on direct adventitious bud induction and plant regeneration of Rosa hybrida Samantha was investigated. The results showed that after an induction period of 8 d on MS medium with 1.5 mg L-1 TDZ and 0.05 mg L-1 NAA in the dark and a subculture on MS medium with 0.5 mg L-1 BA and 0.01 mg L-1 NAA under light, the best plant regeneration was obtained and the regeneration frequencies of leaflets and petioles were 51.8 and 10% respectively. There was no significant difference in regeneration ability between leaflets at different sites of the compound leaves, longer time of induction in the dark or high concentration of auxin would cause callus formation, which was disadvantageous for shoot regeneration, and the regeneration frequency was significantly reduced. This regeneration system could be applied for genetic transformation of this cultivar in the future.展开更多
[Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of Hemerocallis hybrida.[Method] Tender pedicels of dwarf H.hybrida were used as experimental materials to select callu...[Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of Hemerocallis hybrida.[Method] Tender pedicels of dwarf H.hybrida were used as experimental materials to select callus induction,subculture and rooting media for rapid propagation of H.hybrida.[Result] MS+2.0-3.0 mg/L 6-BA+0.2-0.3 mg/L NAA,MS+1.0-2.0 mg/L 6-BA+0.1-0.2 mg/L NAA,MS and 1/2MS+0.2 mg/L NAA were the appropriate medium for callus induction,subculture and rooting,respectively.[Conclusion] The in vitro culture and clustered seedling rooting technology used in this study are effective methods for rapid propagation of H.hybrida,which provide technical reference for industrialized production of H.hybrida.展开更多
As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a ...As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a necessary element for plant growth, and soluble silicon can improve plant resilience. To improve the drought resilience of Petunia hybrida, the silicon transporter protein OsLsi1 and OsLsi2 genes cloned from rice(Oryza sative) were transferred into Petunia hybrida by Agrobacterium-mediated method, and finally got 26 and 32 positive plants, respectively by PCR and RT-PCR detections. With a control of non-transgenic plants, the obtained transgenic plants were taken by drought treatment stress for 0, 4, 7, 10 and 14 days, then re-watered and measured physiological indexes as malondialdehyde(MDA) content, free proline(Pro) content, superoxide dismutase(SOD) activity and peroxidase(POD) activity to study the effect of Petunia's drought resistance. All the results proved that the silicon transporter protein OsLsi1 and OsLsi2 genes were normally transcripted and expressed in transgenic Petunia hybrida; OsLsi1 gene could improve the abilities of plants' drought resistance and recover after drought stress, while OsLsi2 gene could reduce the above abilities. The order of the drought resistance ability of the three strains from strong to weak was OsLsi1>CK>OsLsi2; and silicon indeed improved the ability of drought resistance as well. All these results provided a new way to improve the drought resistance of Petunia, and laid a foundation to improve the ability of garden plants' drought resistance and water saving.展开更多
The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction(RT-PCR)and RACE.The result of sequence analysis indicated that Mlo gene from Pericallis hybrida B.Nord.contained about 1 296 bp ...The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction(RT-PCR)and RACE.The result of sequence analysis indicated that Mlo gene from Pericallis hybrida B.Nord.contained about 1 296 bp open reading frame and encoded 431 amino acids.According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis,Mlo gene shared over 85%nucleotide homology and 98%amino acid homology.Finally,through semi-quantitative-PCR and fluorescence quantitative analysis,we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.展开更多
The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local c...The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local conditions.The experiment included four treatments with three kinds of forliar fertilizers—Dau Trau MK 30-10-5,gibberellin 25-10-10 and abscisic acid.The control treatment(T0)used sterilized water without foliar fertilizer.The results showed that all the foliar fertilizers influenced well on growth and development of Petunia hybrida.Dau Trau MK 30-10-5 helped Petunia hybrida have high quality and high value/cost ratio than the others.展开更多
基金The authors thank Dr.Manzhu Bao(Huazhong Agricultural University,Wuhan,China),Dr.Hibrand-Saint Oyant L.(INRA,Agrocampus-Ouest,Universitéd’Angers,Beaucouzé,France)and Dr.Fabrice Foucher(INRA,78026 Versailles Cedex,France)for their excellent suggestions.We are also grateful to Dr.Wenxue Li and Dr.Hongqiu Wang(Chinese Academy of Agricultural Sci-ences,Beijing,China)for assistance with the experiments.This work was supported by grants from National Natural Science Foundation of China(Grant No.31522049)Construction of Beijing Science and Technology Innovation and Service Capacity in Top Subjects(Grant No.CEFF-PXM2019_014207_000032).
文摘Rose(Rosa hybrida)is widely used for cut flowers and as garden plants.Stable and efficient transformation system is required for functional genomics of rose.Here,we established an efficient transformation method for rose using Agrobacterium tumefaciens-mediated transformation of embryogenic callus.Expanding rose leaves were used as explants to induce somatic embryos,which were subjected to transformation with A.tumefaciens strain GV3101 using Green Fluorescence Protein(GFP)as a marker gene.It took about 8 months to generate transgenic shoots from embryogenic callus.PCR,RT-PCR,Southern and Western blotting,as well as stereoscopic fluorescence microscopy analysis demonstrated that GFP transgenes integrated stably into the rose genome.According to our data,a transformation efficiency of up to 6%can be achieved by following this optimized protocol.
基金the Basic Research Program of Yunnan Province-Youth Project(Grant No.2019FD030)the National Natural Science Foundation of China(Grant No.31960608)the Ten-thousand Talents Program of Yunnan Province–Yunling Scholar of Industrial Technology Leading Talent Project(Grant No.Yun Fagai Renshi[2018]No.212).
文摘Meiotic recombination and the resulting novel allele combinations are fundamental to plant breeding and the identification of superior hybrids.However,the rate of meiotic crossovers is naturally suppressed below its potential ability,which prevents plant breeding efficiency.Nine suppressors of meiotic recombination have been identified in the model plant Arabidopsis and in other crop species.Mutations in these genes can lead to increased recombination frequency and could therefore potentially be used to create hyper-recombinant lines for ornamental breeding.In Gerbera hybrida,the anti-crossover factors remain elusive.In this study,we isolated and cloned TOP3αfrom flower buds of G.hybrida,and it encoded 935 amino acids with three conserved domains TOPRIM,TOP1Ac and zf-GR.Moreover,TOP3αwas the highest expressed at the flower bud stage,which coincided with the occurrence of meiotic recombination,suggesting that TOP3αis associated with the regulation of meiotic recombination in G.hybrida.
基金funded by Yunnan Fundamental Research Projects(Grant No.2019FD030)Major Science and Technology Project of Yunnan Provincial Department of Science and Technology(Grant Nos.2019ZG006,202102AE090052)Ten-thousand Talents Program of Yunnan Province–Yunling Scholar of Industrial Technology Leading Talent Project(Grant No.Yun Fagai Renshi[2018]No.212)。
文摘Gerbera Hybrida is one of the important cut flowers across the world.The novel traits are the primarily market requirements and the breeding targets,mainly determined by the degree of genetic variation after hybridization.However,meiotic recombination is highly conserved in most eukaryotes which suppressed the crossover formation and limited the genetic diversity.Recently,several meiotic recombination suppressors have been identified and characterized in plants,whereas it remains elusive in G.hybrida.In order to characterize the expression patterns of these suppressors in G.hybrida,20 candidate reference genes were identified from the transcriptome datasets of G.hybrida,and their expression stabilities during plant development were evaluated by geNorm,NormFinder and BestKeeper.Although the most stable reference genes were variable in different softwares,comprehensive ranking revealed that PGK2 was the most stable reference gene and GAPDH was the most unstable one.The expression patterns of FANCM,FIGL1,RECQ4,RM1,and FLIP further validated that PGK2 was suitable for normalization of gene expression.Our study identified a reliable reference gene for gene expression during meiotic recombination,and provided functional insights into meiotic recombination suppressors in G.hybrida.
基金Supported by the National Natural Science Foundation of China (Grant No.30170666)
文摘To develop a transformation protocol of Rosa hybrida Samantha via Agrobacterium tumefaciens, the authors examined the effect of different factors on T-DNA transfer by measuring transient expression levels of an intron-containing -glucuronidase gene. The results indicate that explant, light condition, salt concentration and acetosyringone (AS) concentration in co-culture medium are the most important factors, and factors like co-culture temperature, co-culture period and bacteria density have a strong effect on the growth of bacteria and then T-DNA transfer. Optimized co-cultivation was performed by inoculation of embryogenic callus with bacteria at a density of OD600= 0.50.8 for 20 min and co-culture in darkness under 23 C on medium with 1/2 MS salts and 300 mol稬1 AS for 3 d.
基金Supported by Science and Technology Plan Major Projects of Fujian Province(2015NZ0002-1)
文摘In our previous study,a gene predicted to encode a Tyrosine aminotransferase( TAT) was found to be significantly up-regulated in root rot diseased Gerbera by transcriptome sequencing. To confirm the genes and investigate the function,we cloned the gene by RT-PCR and then conduct bioinformatic analyses. In this study,a 1 537 bp long c DNA sequence of this gene( named as Gh TAT) was firstly cloned,which contained a coding region of 1 233 bp,which was predicted to encode a protein of 410 amino acids. Bioinformatic analysis showed that the Gh TAT was a stable hydrophobic protein without signal peptide. Subcellular location prediction result indicated that this protein located in chloroplast,which is the biosynthesis position of tyrosine and the derived products of tyrosine biosynthesis pathway. Moreover,typical Tyrosine aminotransferase domain was found in this protein,indicating that it is a TAT. According to the TAT-based phylogenetic analysis and similarity analysis,the closest relationship and highest similarity was found between Gh TAT and Halianthus annuus TAT,which again verified the TAT property of Gh TAT. Tyrosine aminotransferase( TAT) is the first enzyme in tyrosine biosynthesis pathway,whose products include many antioxidant substances such as tocopherols and tocotrienols. The up-regulation of Gh TAT in root rot diseased gerbera suggests that it may play an important role in response to the root rot pathogen infection. In addition,60 phosphorylation sites( accounting for 14. 6%) were found in this protein,suggesting that the expression of this protein and its encoding gene were greatly influenced by the phosphorylation reactions.
基金the National Natural Science Foundation of China(30170666).
文摘Effect of explant, site of leaflet, induction period in the dark and combinations of plant growth regulators on direct adventitious bud induction and plant regeneration of Rosa hybrida Samantha was investigated. The results showed that after an induction period of 8 d on MS medium with 1.5 mg L-1 TDZ and 0.05 mg L-1 NAA in the dark and a subculture on MS medium with 0.5 mg L-1 BA and 0.01 mg L-1 NAA under light, the best plant regeneration was obtained and the regeneration frequencies of leaflets and petioles were 51.8 and 10% respectively. There was no significant difference in regeneration ability between leaflets at different sites of the compound leaves, longer time of induction in the dark or high concentration of auxin would cause callus formation, which was disadvantageous for shoot regeneration, and the regeneration frequency was significantly reduced. This regeneration system could be applied for genetic transformation of this cultivar in the future.
基金Supported by China Agricultural University (Yantai) Project(yt2007.14)
文摘[Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of Hemerocallis hybrida.[Method] Tender pedicels of dwarf H.hybrida were used as experimental materials to select callus induction,subculture and rooting media for rapid propagation of H.hybrida.[Result] MS+2.0-3.0 mg/L 6-BA+0.2-0.3 mg/L NAA,MS+1.0-2.0 mg/L 6-BA+0.1-0.2 mg/L NAA,MS and 1/2MS+0.2 mg/L NAA were the appropriate medium for callus induction,subculture and rooting,respectively.[Conclusion] The in vitro culture and clustered seedling rooting technology used in this study are effective methods for rapid propagation of H.hybrida,which provide technical reference for industrialized production of H.hybrida.
基金Supported by the Fund of Science and Technology Research Project of Education Department in Heilongjiang Province(12531014)
文摘As one of the important materials in landscaping for flower terrace and border, Petunia hybrida needs high environmental conditions and its growth is seriously influenced by the drought. Silicon is considered to be a necessary element for plant growth, and soluble silicon can improve plant resilience. To improve the drought resilience of Petunia hybrida, the silicon transporter protein OsLsi1 and OsLsi2 genes cloned from rice(Oryza sative) were transferred into Petunia hybrida by Agrobacterium-mediated method, and finally got 26 and 32 positive plants, respectively by PCR and RT-PCR detections. With a control of non-transgenic plants, the obtained transgenic plants were taken by drought treatment stress for 0, 4, 7, 10 and 14 days, then re-watered and measured physiological indexes as malondialdehyde(MDA) content, free proline(Pro) content, superoxide dismutase(SOD) activity and peroxidase(POD) activity to study the effect of Petunia's drought resistance. All the results proved that the silicon transporter protein OsLsi1 and OsLsi2 genes were normally transcripted and expressed in transgenic Petunia hybrida; OsLsi1 gene could improve the abilities of plants' drought resistance and recover after drought stress, while OsLsi2 gene could reduce the above abilities. The order of the drought resistance ability of the three strains from strong to weak was OsLsi1>CK>OsLsi2; and silicon indeed improved the ability of drought resistance as well. All these results provided a new way to improve the drought resistance of Petunia, and laid a foundation to improve the ability of garden plants' drought resistance and water saving.
基金Supported by Postdoctoral Scientifi c Research Foundation of Heilongjiang Province(LBH-Q10144)the Natural Science Foundation of Heilongjiang Province(C201112)Northeast Agricultural University Doctoral Research Fund(200830)
文摘The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction(RT-PCR)and RACE.The result of sequence analysis indicated that Mlo gene from Pericallis hybrida B.Nord.contained about 1 296 bp open reading frame and encoded 431 amino acids.According to the comparison of the exogenous gene sequences by BLAST analysis and phylogenetic analysis,Mlo gene shared over 85%nucleotide homology and 98%amino acid homology.Finally,through semi-quantitative-PCR and fluorescence quantitative analysis,we found that Mlo gene showed the highest expression levels in leaves and the lowest in roots after inoculated with powdery mildew pathogen for different days.
文摘The experiment was conducted in winter-spring 2015-2016 in Thua Thien Hue to identify different foliar fertilizer for Petunia hybrida having good growth and development,beautiful colors and long lifetime under local conditions.The experiment included four treatments with three kinds of forliar fertilizers—Dau Trau MK 30-10-5,gibberellin 25-10-10 and abscisic acid.The control treatment(T0)used sterilized water without foliar fertilizer.The results showed that all the foliar fertilizers influenced well on growth and development of Petunia hybrida.Dau Trau MK 30-10-5 helped Petunia hybrida have high quality and high value/cost ratio than the others.
