The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA P...The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE4(RhSPL4)positively regulates flowering time in rose.Transient silencing or overexpression transgenic rose plants of RhSPL4 exhibited delayed or early flowering,respectively.Analysis of transcriptome data from transgenic lines overexpressing RhSPL4 compared to the wild type indicated that differentially expressed genes were significantly enriched in the circadian rhythm pathway.Among the proteins encoded by these genes,RhSPL4 binds to the promoter of PSEUDO-RESPONSE REGULATOR 5-LIKE(RhPRR5L),as revealed in yeast one-hybrid,dual-Luciferase/Renilla luciferase reporter,chromatin immunoprecipitation-quantitative PCR and electrophoretic mobility shift assay.Furthermore,RhSPL4 specifically binds to the478 to441 bp region of the RhPRR5L promoter and activates its transcription.The silencing of RhPRR5L delayed flowering time in rose,resembling the phenotype of RhSPL4-silenced plants.Together,these results indicate that the RhSPL4-RhPRR5L module positively regulates flowering time in rose,laying the foundation for the genetic improvement of flowering time in this important horticultural crop.展开更多
Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, t...Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, the regulatory interplay between anthocyanin accumulation and tissue acidification processes remains somewhat unclear. The present study characterized an activator Rh MYB114 and a repressor Rh MYB16,which functioned synergistically in anthocyanin accumulation and tissue acidification in rose. Transforming tobacco and roses by overexpression, the introduction of Rh MYB114 resulted in an increase in anthocyanin levels and a noticeable decrease in p H in the petal cells of both rose and tobacco, whereas Rh MYB16 introduction led to inverse effects. To further clarify the underlying the regulatory mechanisms, the yeast two-hybrid(Y2H), bimolecular fluorescence complementation(Bi FC) and dual-luciferase(LUC) were employed. The results showed that Rh MYB16 competed with Rh MYB114, bound to Rhb HLH3 or Rhb HLH33, and inhibited its ability to induce the expression of genes related to anthocyanin biosynthesis and acidification. Our findings revealed a feedback mechanism for the regulation of anthocyanin synthesis and tissue acidification involving Rh MYB114, which stimulated the transcriptional expression of Rh MYB16, whose encoded protein Rh MYB16, in turn, negatively regulated the transcriptional expression of Rh MYB114. Therefore, this study underscores the pivotal roles of the Rh MYB114-Rh MYB16 loop in regulating anthocyanin synthesis and tissue acidification, offering insights into metabolic manipulation to enhance the aesthetic appeal of roses.展开更多
To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids...To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids were made into fungal elicitors.These fungal elicitors were added into the DE medium with concentrations of 40,60 and 80 ml/L,respectively.After a 90-d culturing,the effects of fungal elicitors on the growth of D.hybrida cultivar ‘088＇ tissue culture seedlings were studied.The results showed that treatment 13（T13） extremely significantly increased the fresh weight,but other treatment groups had no significant effects.In addition,T1,T5,T9,T11 and T13 extremely significantly influenced the contents of chlorophyll a and chlorophyll a ＋ b.However,T1 and T11 had extremely significantly effect on the content of chlorophyll b.Combining the effects on fresh weight and chlorophyll content,it could be concluded T13（40 ml/L of Y05） has promoting effects on the growth of D.hybrida tissue culture seedlings.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
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
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
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
[ Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of HemerocaUis hybrida. [ Method] Tender pedicels of dwarf H. hybrida were used as experimental materials to sdect cal...[ Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of HemerocaUis hybrida. [ Method] Tender pedicels of dwarf H. hybrida were used as experimental materials to sdect 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 me- dium 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 technieal reference for industrialized production of H. hybrida.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
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 M/o gene from Pericallis hybrida B. Nord. contained about 12...The full-length Mlo gene was obtained by reverse transcription polymerase chain reaction （RT-PCR） and RACE. The result of sequence analysis indicated that M/o gene from Pericallis hybrida B. Nord. contained about 1296bp 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
The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process ...The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production and 1- aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’ than control during vasing. cDNA fragments of three Rh-ACSs and one Rh- ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-in- duced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhib-ited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in ‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding.展开更多
A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method medi...A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method mediated by Agrobacterium. A series of novel phenotypes was observed in transgenic petunia plants, including the formation of ectopic spikes on the adaxial surface of corollas and small petals on the abaxial surface of corollas, fusion of floral organs, shortening of corolla midribs, the formation of tumor-like knots along the midrib on the abaxial surface and serrated lobs of corolla margins, and alterations in petal color; except for changes in the leaves and plant architecture, RT-PCR showed that the Pttknl gene was expressed in the leaves of different petunia transgenic plants, whereas no signal was detected in wild-type plants. The possible function of Pttknl in leaf and flower development is discussed.展开更多
基金supported by Yunnan Province Agricultural Joint Key Project(Grant No.202401BD070001-016)the National Natural Science Foundation of China(Grant No.32202530)+3 种基金Talent Introduction and Training Project of Yunnan Academy of Agricultural Sciences(Grant No.2024RCYP-09)Fundamental Research Project(Grant No.202401CF070046)Xingdian Talent support program(XDYC-QNRC-2023-0457)Yunnan Technology Innovation Center of Flower Technique.
文摘The proper flowering time of rose(Rosa hybrida)is vital for the market value of this horticultural crop,but the mechanism regulating this trait is largely unclear.Here,we found that the transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE4(RhSPL4)positively regulates flowering time in rose.Transient silencing or overexpression transgenic rose plants of RhSPL4 exhibited delayed or early flowering,respectively.Analysis of transcriptome data from transgenic lines overexpressing RhSPL4 compared to the wild type indicated that differentially expressed genes were significantly enriched in the circadian rhythm pathway.Among the proteins encoded by these genes,RhSPL4 binds to the promoter of PSEUDO-RESPONSE REGULATOR 5-LIKE(RhPRR5L),as revealed in yeast one-hybrid,dual-Luciferase/Renilla luciferase reporter,chromatin immunoprecipitation-quantitative PCR and electrophoretic mobility shift assay.Furthermore,RhSPL4 specifically binds to the478 to441 bp region of the RhPRR5L promoter and activates its transcription.The silencing of RhPRR5L delayed flowering time in rose,resembling the phenotype of RhSPL4-silenced plants.Together,these results indicate that the RhSPL4-RhPRR5L module positively regulates flowering time in rose,laying the foundation for the genetic improvement of flowering time in this important horticultural crop.
基金supported by the National Natural Science Foundation of China(Grant No.32171851)Natural Science Foundation of Zhejiang(Grant No.LZ23C160003)the Fundamental Research Funds for the Zhejiang A&F University(Grant No.2016FR033)。
文摘Coloration in rose(Rosa hybrida) petals is primarily determined by anthocyanin accumulation in vacuoles, and vacuolar acidification plays a central role in controlling the accumulation of this pigment. Nevertheless, the regulatory interplay between anthocyanin accumulation and tissue acidification processes remains somewhat unclear. The present study characterized an activator Rh MYB114 and a repressor Rh MYB16,which functioned synergistically in anthocyanin accumulation and tissue acidification in rose. Transforming tobacco and roses by overexpression, the introduction of Rh MYB114 resulted in an increase in anthocyanin levels and a noticeable decrease in p H in the petal cells of both rose and tobacco, whereas Rh MYB16 introduction led to inverse effects. To further clarify the underlying the regulatory mechanisms, the yeast two-hybrid(Y2H), bimolecular fluorescence complementation(Bi FC) and dual-luciferase(LUC) were employed. The results showed that Rh MYB16 competed with Rh MYB114, bound to Rhb HLH3 or Rhb HLH33, and inhibited its ability to induce the expression of genes related to anthocyanin biosynthesis and acidification. Our findings revealed a feedback mechanism for the regulation of anthocyanin synthesis and tissue acidification involving Rh MYB114, which stimulated the transcriptional expression of Rh MYB16, whose encoded protein Rh MYB16, in turn, negatively regulated the transcriptional expression of Rh MYB114. Therefore, this study underscores the pivotal roles of the Rh MYB114-Rh MYB16 loop in regulating anthocyanin synthesis and tissue acidification, offering insights into metabolic manipulation to enhance the aesthetic appeal of roses.
基金Supported by National Nonprofit Institute Research Grant of CATAS-TCGRI(1630032014027)Special Fund for Agro-scientific Research in the Public Interest(201203071)Special Project for Scientific and Technological Achievements Demonstration and Promotion in Hainan Province(SQ2013CGSF0006)~~
文摘To optimize the technique of rapid propagation of Dendrobium hybrida seedlings and to explore a hormone-free tissue culture method for D.hybrida,six kinds of mycorrhizal fungi which were isolated from the wild orchids were made into fungal elicitors.These fungal elicitors were added into the DE medium with concentrations of 40,60 and 80 ml/L,respectively.After a 90-d culturing,the effects of fungal elicitors on the growth of D.hybrida cultivar ‘088＇ tissue culture seedlings were studied.The results showed that treatment 13（T13） extremely significantly increased the fresh weight,but other treatment groups had no significant effects.In addition,T1,T5,T9,T11 and T13 extremely significantly influenced the contents of chlorophyll a and chlorophyll a ＋ b.However,T1 and T11 had extremely significantly effect on the content of chlorophyll b.Combining the effects on fresh weight and chlorophyll content,it could be concluded T13（40 ml/L of Y05） has promoting effects on the growth of D.hybrida tissue culture seedlings.
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 HemerocaUis hybrida. [ Method] Tender pedicels of dwarf H. hybrida were used as experimental materials to sdect 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 me- dium 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 technieal reference for industrialized production of H. hybrida.
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 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 M/o gene from Pericallis hybrida B. Nord. contained about 1296bp 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.
基金This work was supported by the Nat ional Natural Science Foundation of China(Grant No.30471220).
文摘The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, ‘Samantha’, whose opening process is promoted, and ‘Kardinal’, whose opening process is inhibited by ethylene. Ethylene production and 1- aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in ‘Samantha’, and they were much more dramatically enhanced and peaked at the later stage (stage 4) in ‘Kardinal’ than control during vasing. cDNA fragments of three Rh-ACSs and one Rh- ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-in- duced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in ‘Kardinal’ than that of ‘Samantha’. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhib-ited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in ‘Kardinal’. Our results suggests that ‘Kardinal’ is more sensitive to ethylene than ‘Samantha’; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in ‘Samantha’ and the inhibition in ‘Kardinal’. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding.
基金Supported by the Gansu Provincial Natural Science Foundation of China （ZS021-A25-047-N） and Xi＇an Urban Rural Construction Committee.Acknowledgements The authors thank Dr. 0lof 0lsson （Goeteberg University, Goeteberg, Sweden） for providing the plasmid containing 35S：：Pttkn1.
文摘A novel knottedl-like homeobox （knox） gene, Pttknl （Populus tremulaxtremuloides knottedl）, isolated from the cambial region of hybrid aspen, was introduced into Petunia hybrida Vilm. using the leafdisc method mediated by Agrobacterium. A series of novel phenotypes was observed in transgenic petunia plants, including the formation of ectopic spikes on the adaxial surface of corollas and small petals on the abaxial surface of corollas, fusion of floral organs, shortening of corolla midribs, the formation of tumor-like knots along the midrib on the abaxial surface and serrated lobs of corolla margins, and alterations in petal color; except for changes in the leaves and plant architecture, RT-PCR showed that the Pttknl gene was expressed in the leaves of different petunia transgenic plants, whereas no signal was detected in wild-type plants. The possible function of Pttknl in leaf and flower development is discussed.
