Tissue culture has been widely used for mass propagation of Phalaenopsis. However, somaclonal variation occurred during micropropagation process posed a severe problem by affecting product quality. In this study, wild...Tissue culture has been widely used for mass propagation of Phalaenopsis. However, somaclonal variation occurred during micropropagation process posed a severe problem by affecting product quality. In this study, wild type and peloric flower buds of Phalaenopsis hybrids derived from flower stalk nodal culture were used for cDNA-RAPD and cDNA suppression subtractive hybridization analyses in order to study their genetic difference in terms of expressed sequence tags. A total of 209 ESTs from normal flower buds and 230 from mutants were sequenced. These ESTs sequences can be grouped into several functional categories involved in different cellular processes including metabolism, signal transduction, transcription, cell growth and division, protein synthesis, and protein localization, and into a subcat- egory of proteins with unknown function. Cymbidium mosaic virus transcript was surprisingly found expressed fre- quently in the peloric mutant of P. Little Mary. Real-time RT-PCR analysis on selected ESTs showed that in mutant flower buds, a bZIP transcription factor (TGA1a-like protein) was down-regulated, while up-regulated genes include auxin-regulated protein kinase, cyclophilin, and TCP-like genes. A retroelement clone was also preferentially expressed in the peloric mutant flowers. On the other hand, ESTs involved in DNA methylation, chromatin remodeling and post- transcriptional regulation, such as DNA methyltransferase, histone acetyltransferase, ERECTA, and DEAD/DEAH RNA helicase, were enriched in normal flower buds than the mutants. The enriched transcripts in the wild type indicate the down regulation of these transcripts in the mutants, and vice versa. The potential roles of the analyzed transcripts in the development of Phalaenopsis flowers are discussed.展开更多
This paper studied the rapid propagation technology of Phalaenopsis hybrid by using the peduncle buds as the explants, and screened out a kind of culture medium, which was simple, rapid, available and high rate of pro...This paper studied the rapid propagation technology of Phalaenopsis hybrid by using the peduncle buds as the explants, and screened out a kind of culture medium, which was simple, rapid, available and high rate of propagation.展开更多
Stress response at the protein level to viral infection in orchid plants has not been extensively investigated to date. To understand the proteomic basis of Phalaenopsis amabilis’s responses to Cymbidium Mosaic virus...Stress response at the protein level to viral infection in orchid plants has not been extensively investigated to date. To understand the proteomic basis of Phalaenopsis amabilis’s responses to Cymbidium Mosaic virus (CymMV), and/or Odontoglossum ring spot virus (ORSV), the total proteins were extracted from Phalaenopsis amabilis leaves infected with CymMV, ORSV, or both respectively. Differentially expressed proteins were identified by two-dimensional electrophoresis, and 27 of these proteins that had significant changes were further examined by mass spectrometry. Comparing CymMV-infected leaves with mock-inoculated ones, 2 proteins were significantly up-regulated, 9 were significantly down-regulated and 1 previously undetected protein was identified. 10 proteins were significantly up-regulated, 3 significantly down-regulated and 1 previously undetected protein was identified in ORSV-infected leaves. 6 proteins were significantly up-regulated and 9 significantly down-regulated proteins were found in co-infected leaves. These identified proteins are involved in disease resistance, stress response, transcriptional regulation, energy metabolism, protein modification and the previously unknown proteins were not involved with known protein pathways. Proteins significantly up-regulated were ATP sulfurylase, down-regulated proteins included glutamate decarboxylase isozyme 2, RNA polymerase alpha subunit and chloroplastic peptide deformylase 1A were proteins with similar alteration trend after all infection treatments. Significantly up-regulated were Thioredoxin H-type and down-regulated Cytosolic phosphoglycerate kinase I which were proteins that have been shown to be specifically regulated by the infection with CymMV. Significantly up-regulated were proteins like Rubisco large subunit, Triosephosphate isomerase, NADP-specific isocitrate dehydrogenase and Cinnamoyl CoA reductase CCR2 by the infection of ORSV. Protein regulation in coinfected leaves followed a pattern similar to that of any of the single virus infection results.展开更多
The Phalaenopsis aphrodite industry has the characteristics of high technology content,industrialization,modern management,organic combination of industry and strong radiation effect. On the basis of investigation,we ...The Phalaenopsis aphrodite industry has the characteristics of high technology content,industrialization,modern management,organic combination of industry and strong radiation effect. On the basis of investigation,we expounded the current development situation of the P. aphrodite industry in Guangdong,analyzed the aspects of production elements and demand conditions based on the " diamond theory",and put forward suggestion and countermeasures for the development of the P. aphrodite industry.展开更多
Phalaenopsis amabilis industry in Zhangzhou City is analyzed by SWOT method,and the strengths,weaknesses,opportunities and threats of P.amabilis industry in the city are expounded.Furthermore,some suggestions for the ...Phalaenopsis amabilis industry in Zhangzhou City is analyzed by SWOT method,and the strengths,weaknesses,opportunities and threats of P.amabilis industry in the city are expounded.Furthermore,some suggestions for the development of P.amabilis industry in Zhangzhou City are put forward,in order to provide certain references for the development of P.amabilis industry.展开更多
A key step in regulating the flowering of Phalaenopsis is to control the emergence of the flower stalk or spiking. Light quality is an important factor to regulate plant reproduction. We used either red or blue light ...A key step in regulating the flowering of Phalaenopsis is to control the emergence of the flower stalk or spiking. Light quality is an important factor to regulate plant reproduction. We used either red or blue light emitting diodes (LEDs) to substitute for 10% photosynthetic photon flux emitted by white fluorescent tubes as the control (WC) at approximately 70 μmole·m<sup>-2</sup>·s<sup>-1</sup> to examine the effects of enhanced red (WR) or blue (WB) light on spiking and the dawn-to-dusk fluctuations in the photosynthetic pigments and carbohydrates of Phalaenopsis aphrodite, which were grown in 7.5 cm diameter pots for six weeks. WR treatment significantly elevated the ratio of red to far-red light and the level of phytochrome photostationary state in concurrence with an increase in the spiking ratio and length, but had little effect on the concentrations of chlorophyll a and chlorophyll b in the leaf, levels of soluble sugars such as glucose, fructose, sucrose, and insoluble starch in the leaf and shortened stem, the locus of spiking, when compared to the other two treatments. Evidently, the spiking of Phalaenopsis is improved by dependent on the relative amount of active phytochrome expressed in the photostationary state.展开更多
To study the main aroma components of Phalaenopsis orchid and their relationship with colors,10 fragrant cultivars with different colors,like pink,rose,yellow,and purple,were used as samples in this experiment.Headspa...To study the main aroma components of Phalaenopsis orchid and their relationship with colors,10 fragrant cultivars with different colors,like pink,rose,yellow,and purple,were used as samples in this experiment.Headspace-gas chromatography-mass spectrometry was used to determine the main components of floral fragrance and analyze the correlation between floral color and fragrance.The results showed that the main aroma components of the 10 fragrant cultivars of Phalaenopsis were alcohols,alkenes,esters,and benzene ring compounds,and the main aroma components of different cultivars were diverse.The main aroma components of yellow fragrant flowers were esters,alcohols,and alkenes.The purple and pink series were alcohols and phenyl rings.There was a certain correlation between flower color and floral fragrance.There was a significant positive correlation between esters and flower color C^(*)value,and a significant negative correlation between alkenes and flower color h value.There was a significant negative correlation between alcohol and flower color C^(*)value,and a significant positive correlation between alcohol and L^(*)value.The content of benzene compounds was negatively correlated with L^(*)and positively correlated with h value.This may be related to the synthesizing of anthocyanins and benzene ring compounds through the phenylpropanoid metabolic pathway.In this paper,the correlation between Phalaenopsis floral color and fragrance was studied,and the synthetic pathway of floral color and fragrance components was analyzed.The proposed method and research data can provide a theoretical basis for floral color breeding and fragrance synthesis.展开更多
Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on ...Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on the early stage of browning in explants of the orchid Phalaenopsis. Our results show that PPO activity was significantly higher in explants cultured for 3 d than in the 0 h control. The levels of PPO transcripts and PPO protein were significantly higher in explants cultured for 6 h compared to the 0 h control; these high expression levels were maintained over increasing cultivation time. Cycloheximide(CHX) treatment reduced PPO transcript levels, PPO protein levels, and PPO enzyme activity. High levels of PPO m RNA and PPO protein were detected in the cytoplasm and vascular bundles of Phalaenopsis explants cultured for 6 h compared to explants cultured for 0 h, 24 h, and 3 d. CHX treatment did not significantly affect the distribution of PPO m RNA and PPO protein in explant tissues, but their levels were significantly lower than those of the untreated control.展开更多
Identifying the species composition of a microbial ecosystem is often hampered by difficulties in culturing the organisms and in the low sequencing depth of traditional DNA barcoding.Metagenomic analysis,a huge-scale ...Identifying the species composition of a microbial ecosystem is often hampered by difficulties in culturing the organisms and in the low sequencing depth of traditional DNA barcoding.Metagenomic analysis,a huge-scale nucleotide-sequence-based tool,can overcome such difficulties.In this study,Sanger sequencing of 500 nrITS clones uncovered 29 taxa of 19 fungal genera,whereas metagenomics with next-generation sequencing identified 512 operational taxonomic units(OTUs)for ITS1/2 and 364 for ITS3/4.Nevertheless,high throughput sequencing of PCR amplicons of ITS1/2,ITS3/4,nrLSU-LR,nrLSU-U,mtLSU,and mtATP6,all with at least 1,300×coverage and about 21 million reads in total,yielded a very diverse fungal composition.The fact that 74%of the OTUs were exclusively uncovered with single barcodes indicated that each marker provided its own insights into the fungal flora.To deal with the high heterogeneity in the data and to integrate the information on species composition across barcodes,a rank-scoring strategy was developed.Accordingly,205 genera among 64 orders of fungi were identified in healthy Phalaenopsis roots.Of the barcodes utilized,ITS1/2,ITS3/4,and nrLSU-U were the most competent in uncovering the fungal diversity.These barcodes,though detecting different compositions likely due to primer preference,provided complementary and comprehensive power in deciphering the microbial diversity,especially in revealing rare species.展开更多
In the attempt to discover new genes involved in the floral development in monocotyledonousin species,we have cloned and characterized the homologous PISTALLATA-like(PI-like)gene from Phalaenopsis hybrid cultivar name...In the attempt to discover new genes involved in the floral development in monocotyledonousin species,we have cloned and characterized the homologous PISTALLATA-like(PI-like)gene from Phalaenopsis hybrid cultivar named PhPI9(Phalaenopsis PI STILLATA#9).The cDNA of PhPI9 has a fragment of 834 bp and has 60%identity with the PISTILATA from Arabidopsis.The deduced amino acid sequence of PhPI9 had the typical PI-motif.It also formed a subclade with other monocot PI-type genes in phylogenetic analysis.Southern analysis showed that PhPI9 was present in the Phalaenopsis orchid genome as a single copy.Furthermore,it was expressed only in the lip of the Phalaenopsis flower and no expression was detected in vegetative organs.Thus,as a B-function MADS-box gene,PhPI9 specifies floral organ identity in orchids.展开更多
The molecular basis of orchid flower development involves a specific regulatory program in which MADS-box transcription factors play a central role.The recent‘perianth code’model hypothesizes that two types of highe...The molecular basis of orchid flower development involves a specific regulatory program in which MADS-box transcription factors play a central role.The recent‘perianth code’model hypothesizes that two types of higher-order heterotetrameric complexes,namely SP complex and L complex,play pivotal roles in the orchid perianth organ formation.Therefore,we explored their roles and searched for other components of the regulatory network.Through the combined analysis for transposase-accessible chromatin with high-throughput sequencing and RNA sequencing of the lip-like petal and lip from Phalaenopsis equestris var.trilip,transcription factor-(TF)genes involved in lip development were revealed.PeNAC67 encoding a NAC-type TF and PeSCL23 encoding a GRAS-type TF were differentially expressed between the lip-like petal and the lip.PeNAC67 interacted with and stabilized PeMADS3,which positively regulated the development of lip-like petal to lip.PeSCL23 and PeNAC67 competitively bound with PeKAN2 and positively regulated the development of lip-like petal to petal by affecting the level of PeMADS3.PeKAN2 as an important TF that interacts with PeMADS3 and PeMADS9 can promote lip development.These results extend the‘perianth code’model and shed light on the complex regulation of orchid flower development.展开更多
文摘Tissue culture has been widely used for mass propagation of Phalaenopsis. However, somaclonal variation occurred during micropropagation process posed a severe problem by affecting product quality. In this study, wild type and peloric flower buds of Phalaenopsis hybrids derived from flower stalk nodal culture were used for cDNA-RAPD and cDNA suppression subtractive hybridization analyses in order to study their genetic difference in terms of expressed sequence tags. A total of 209 ESTs from normal flower buds and 230 from mutants were sequenced. These ESTs sequences can be grouped into several functional categories involved in different cellular processes including metabolism, signal transduction, transcription, cell growth and division, protein synthesis, and protein localization, and into a subcat- egory of proteins with unknown function. Cymbidium mosaic virus transcript was surprisingly found expressed fre- quently in the peloric mutant of P. Little Mary. Real-time RT-PCR analysis on selected ESTs showed that in mutant flower buds, a bZIP transcription factor (TGA1a-like protein) was down-regulated, while up-regulated genes include auxin-regulated protein kinase, cyclophilin, and TCP-like genes. A retroelement clone was also preferentially expressed in the peloric mutant flowers. On the other hand, ESTs involved in DNA methylation, chromatin remodeling and post- transcriptional regulation, such as DNA methyltransferase, histone acetyltransferase, ERECTA, and DEAD/DEAH RNA helicase, were enriched in normal flower buds than the mutants. The enriched transcripts in the wild type indicate the down regulation of these transcripts in the mutants, and vice versa. The potential roles of the analyzed transcripts in the development of Phalaenopsis flowers are discussed.
文摘This paper studied the rapid propagation technology of Phalaenopsis hybrid by using the peduncle buds as the explants, and screened out a kind of culture medium, which was simple, rapid, available and high rate of propagation.
文摘Stress response at the protein level to viral infection in orchid plants has not been extensively investigated to date. To understand the proteomic basis of Phalaenopsis amabilis’s responses to Cymbidium Mosaic virus (CymMV), and/or Odontoglossum ring spot virus (ORSV), the total proteins were extracted from Phalaenopsis amabilis leaves infected with CymMV, ORSV, or both respectively. Differentially expressed proteins were identified by two-dimensional electrophoresis, and 27 of these proteins that had significant changes were further examined by mass spectrometry. Comparing CymMV-infected leaves with mock-inoculated ones, 2 proteins were significantly up-regulated, 9 were significantly down-regulated and 1 previously undetected protein was identified. 10 proteins were significantly up-regulated, 3 significantly down-regulated and 1 previously undetected protein was identified in ORSV-infected leaves. 6 proteins were significantly up-regulated and 9 significantly down-regulated proteins were found in co-infected leaves. These identified proteins are involved in disease resistance, stress response, transcriptional regulation, energy metabolism, protein modification and the previously unknown proteins were not involved with known protein pathways. Proteins significantly up-regulated were ATP sulfurylase, down-regulated proteins included glutamate decarboxylase isozyme 2, RNA polymerase alpha subunit and chloroplastic peptide deformylase 1A were proteins with similar alteration trend after all infection treatments. Significantly up-regulated were Thioredoxin H-type and down-regulated Cytosolic phosphoglycerate kinase I which were proteins that have been shown to be specifically regulated by the infection with CymMV. Significantly up-regulated were proteins like Rubisco large subunit, Triosephosphate isomerase, NADP-specific isocitrate dehydrogenase and Cinnamoyl CoA reductase CCR2 by the infection of ORSV. Protein regulation in coinfected leaves followed a pattern similar to that of any of the single virus infection results.
基金Supported by Huizhou Engineering Vocational College Project (No.2019kt003)。
文摘The Phalaenopsis aphrodite industry has the characteristics of high technology content,industrialization,modern management,organic combination of industry and strong radiation effect. On the basis of investigation,we expounded the current development situation of the P. aphrodite industry in Guangdong,analyzed the aspects of production elements and demand conditions based on the " diamond theory",and put forward suggestion and countermeasures for the development of the P. aphrodite industry.
基金Supported by Fujian Spark Project"Standardized Production and Demonstration of Phalaenopsis amabilis Export Seedlings in Zhangpu County"(2020S0045).
文摘Phalaenopsis amabilis industry in Zhangzhou City is analyzed by SWOT method,and the strengths,weaknesses,opportunities and threats of P.amabilis industry in the city are expounded.Furthermore,some suggestions for the development of P.amabilis industry in Zhangzhou City are put forward,in order to provide certain references for the development of P.amabilis industry.
文摘A key step in regulating the flowering of Phalaenopsis is to control the emergence of the flower stalk or spiking. Light quality is an important factor to regulate plant reproduction. We used either red or blue light emitting diodes (LEDs) to substitute for 10% photosynthetic photon flux emitted by white fluorescent tubes as the control (WC) at approximately 70 μmole·m<sup>-2</sup>·s<sup>-1</sup> to examine the effects of enhanced red (WR) or blue (WB) light on spiking and the dawn-to-dusk fluctuations in the photosynthetic pigments and carbohydrates of Phalaenopsis aphrodite, which were grown in 7.5 cm diameter pots for six weeks. WR treatment significantly elevated the ratio of red to far-red light and the level of phytochrome photostationary state in concurrence with an increase in the spiking ratio and length, but had little effect on the concentrations of chlorophyll a and chlorophyll b in the leaf, levels of soluble sugars such as glucose, fructose, sucrose, and insoluble starch in the leaf and shortened stem, the locus of spiking, when compared to the other two treatments. Evidently, the spiking of Phalaenopsis is improved by dependent on the relative amount of active phytochrome expressed in the photostationary state.
基金supported by the Shandong Province Key Research and Development Plan Project(ID Numbers 2024LZGC026 and 2021LZGC019)Shanghai Science and Technology Agriculture Project(ID Number 2020-02-08-00-12-F01463).
文摘To study the main aroma components of Phalaenopsis orchid and their relationship with colors,10 fragrant cultivars with different colors,like pink,rose,yellow,and purple,were used as samples in this experiment.Headspace-gas chromatography-mass spectrometry was used to determine the main components of floral fragrance and analyze the correlation between floral color and fragrance.The results showed that the main aroma components of the 10 fragrant cultivars of Phalaenopsis were alcohols,alkenes,esters,and benzene ring compounds,and the main aroma components of different cultivars were diverse.The main aroma components of yellow fragrant flowers were esters,alcohols,and alkenes.The purple and pink series were alcohols and phenyl rings.There was a certain correlation between flower color and floral fragrance.There was a significant positive correlation between esters and flower color C^(*)value,and a significant negative correlation between alkenes and flower color h value.There was a significant negative correlation between alcohol and flower color C^(*)value,and a significant positive correlation between alcohol and L^(*)value.The content of benzene compounds was negatively correlated with L^(*)and positively correlated with h value.This may be related to the synthesizing of anthocyanins and benzene ring compounds through the phenylpropanoid metabolic pathway.In this paper,the correlation between Phalaenopsis floral color and fragrance was studied,and the synthetic pathway of floral color and fragrance components was analyzed.The proposed method and research data can provide a theoretical basis for floral color breeding and fragrance synthesis.
基金supported by Natural Science Foundation in China(31070618)
文摘Explant browning is one of the major problems in the tissue culture process, and polyphenol oxidase(PPO), is the major proteases involved in plant tissue browning. We investigated the effects of polyphenol oxidase on the early stage of browning in explants of the orchid Phalaenopsis. Our results show that PPO activity was significantly higher in explants cultured for 3 d than in the 0 h control. The levels of PPO transcripts and PPO protein were significantly higher in explants cultured for 6 h compared to the 0 h control; these high expression levels were maintained over increasing cultivation time. Cycloheximide(CHX) treatment reduced PPO transcript levels, PPO protein levels, and PPO enzyme activity. High levels of PPO m RNA and PPO protein were detected in the cytoplasm and vascular bundles of Phalaenopsis explants cultured for 6 h compared to explants cultured for 0 h, 24 h, and 3 d. CHX treatment did not significantly affect the distribution of PPO m RNA and PPO protein in explant tissues, but their levels were significantly lower than those of the untreated control.
基金This study was financially supported by the National Cheng Kung University and the National Science Council,Taiwan.
文摘Identifying the species composition of a microbial ecosystem is often hampered by difficulties in culturing the organisms and in the low sequencing depth of traditional DNA barcoding.Metagenomic analysis,a huge-scale nucleotide-sequence-based tool,can overcome such difficulties.In this study,Sanger sequencing of 500 nrITS clones uncovered 29 taxa of 19 fungal genera,whereas metagenomics with next-generation sequencing identified 512 operational taxonomic units(OTUs)for ITS1/2 and 364 for ITS3/4.Nevertheless,high throughput sequencing of PCR amplicons of ITS1/2,ITS3/4,nrLSU-LR,nrLSU-U,mtLSU,and mtATP6,all with at least 1,300×coverage and about 21 million reads in total,yielded a very diverse fungal composition.The fact that 74%of the OTUs were exclusively uncovered with single barcodes indicated that each marker provided its own insights into the fungal flora.To deal with the high heterogeneity in the data and to integrate the information on species composition across barcodes,a rank-scoring strategy was developed.Accordingly,205 genera among 64 orders of fungi were identified in healthy Phalaenopsis roots.Of the barcodes utilized,ITS1/2,ITS3/4,and nrLSU-U were the most competent in uncovering the fungal diversity.These barcodes,though detecting different compositions likely due to primer preference,provided complementary and comprehensive power in deciphering the microbial diversity,especially in revealing rare species.
文摘In the attempt to discover new genes involved in the floral development in monocotyledonousin species,we have cloned and characterized the homologous PISTALLATA-like(PI-like)gene from Phalaenopsis hybrid cultivar named PhPI9(Phalaenopsis PI STILLATA#9).The cDNA of PhPI9 has a fragment of 834 bp and has 60%identity with the PISTILATA from Arabidopsis.The deduced amino acid sequence of PhPI9 had the typical PI-motif.It also formed a subclade with other monocot PI-type genes in phylogenetic analysis.Southern analysis showed that PhPI9 was present in the Phalaenopsis orchid genome as a single copy.Furthermore,it was expressed only in the lip of the Phalaenopsis flower and no expression was detected in vegetative organs.Thus,as a B-function MADS-box gene,PhPI9 specifies floral organ identity in orchids.
基金funded by National Key Research and Development Program(grant 2023YFD10005002018YFD000400)Shanghai Engineering Research Center of Plant Germplasm Resources(grant number 17DZ2252700).
文摘The molecular basis of orchid flower development involves a specific regulatory program in which MADS-box transcription factors play a central role.The recent‘perianth code’model hypothesizes that two types of higher-order heterotetrameric complexes,namely SP complex and L complex,play pivotal roles in the orchid perianth organ formation.Therefore,we explored their roles and searched for other components of the regulatory network.Through the combined analysis for transposase-accessible chromatin with high-throughput sequencing and RNA sequencing of the lip-like petal and lip from Phalaenopsis equestris var.trilip,transcription factor-(TF)genes involved in lip development were revealed.PeNAC67 encoding a NAC-type TF and PeSCL23 encoding a GRAS-type TF were differentially expressed between the lip-like petal and the lip.PeNAC67 interacted with and stabilized PeMADS3,which positively regulated the development of lip-like petal to lip.PeSCL23 and PeNAC67 competitively bound with PeKAN2 and positively regulated the development of lip-like petal to petal by affecting the level of PeMADS3.PeKAN2 as an important TF that interacts with PeMADS3 and PeMADS9 can promote lip development.These results extend the‘perianth code’model and shed light on the complex regulation of orchid flower development.
