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1.
Planta ; 259(4): 84, 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38448635

RESUMO

MAIN CONCLUSION: A novel electroporation method for genome editing was performed using plant tissue samples by direct RNPs-introduction in carnation. Genome editing is becoming a very useful tool in plant breeding. In this study, a novel electroporation method was performed for genome editing using plant tissue samples. The objective was to create a flower color mutant using the pink-flowered carnation 'Kane Ainou 1-go'. For this purpose, a ribonucleoprotein consisting of guide RNA and clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) was introduced into the stem tissue to induce mutations in the anthocyanidin synthase (ANS) gene, which is involved in anthocyanin biosynthesis. As the ANS of 'Kane Ainou 1-go' has not been previously isolated, we initially isolated the ANS gene from 'Kane Ainou 1-go' for characterization. Southern hybridization analysis confirmed that the ANS gene was present in the genome as a two-allele gene with a pair of homologous sequences (ANS-1 and 2); these sequences were used as the target for genome editing. Genome editing was performed by introducing #2_single-guide RNA into the stem tissue using the ribonucleoprotein. This molecule was used because it exhibited the highest efficiency in an analysis of cleavage activity against the target sequence in vitro. Cleaved amplified polymorphic sequence analysis of genomic DNA extracted from 85 regenerated individuals after genome editing was performed. The results indicated that mutations in the ANS gene may have been introduced into two lines. Cloning of the ANS gene in these two lines confirmed the introduction of a single nucleotide substitution mutation for ANS-1 in both lines, and a single amino acid substitution in one line. We discussed the possibility of color change by the amino acid substitution, and also the future applications of this technology.


Assuntos
Dianthus , Oxigenases , Humanos , Edição de Genes , RNA Guia de Sistemas CRISPR-Cas , Melhoramento Vegetal , Eletroporação , Ribonucleoproteínas
2.
Plants (Basel) ; 12(17)2023 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-37687366

RESUMO

Chrysanthemum is one of the most commercially used ornamental flowering plants in the world. As chrysanthemum is self-incompatible, the propagation of identical varieties is carried out through cuttings rather than through seed. Axillary bud development can be controlled by changing the temperature; for instance, axillary bud development in some varieties is suppressed at high temperatures. In this study, we focused on the simultaneous axillary bud growth from multiple lines of chrysanthemum upon changing conditions from low to normal temperature. Transcriptome analysis was conducted on the Chrysanthemum morifolium cultivar 'Jinba' to identify the important genes for axillary bud development seen when moved from low-temperature treatment to normal cultivation temperature. We performed RNA-Seq analysis on plants after cold conditions in two-day time-course experiments. Under these settings, we constructed a transcriptome of 415,923 C. morifolium and extracted 7357 differentially expressed genes. Our understanding of Arabidopsis axillary meristem development and growth showed that at least 101 genes in our dataset were homologous to transcription factors involved in the biological process. In addition, six genes exhibited statistically significant variations in expression throughout conditions. We hypothesized that these genes were involved in the formation of axillary buds in C. morifolium after cold conditions.

3.
Plants (Basel) ; 12(13)2023 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-37447127

RESUMO

Chrysanthemum is one of the most commercially important ornamental plants globally, of which many new varieties are produced annually. Among these new varieties, many are the result of crossbreeding, while some are the result of mutation breeding. Recent advances in gene and genome sequencing technology have raised expectations about the use of biotechnology and genome breeding to efficiently breed new varieties. However, some features of chrysanthemum complicate molecular biological analysis. For example, chrysanthemum is a hexaploid hyperploid plant with a large genome, while its genome is heterogeneous because of the difficulty of obtaining pure lines due to self-incompatibility. Despite these difficulties, an increased number of reports on transcriptome analysis in chrysanthemum have been published as a result of recent technological advances in gene sequencing, which should deepen our understanding of the properties of these plants. In this review, we discuss recent studies using gene engineering, genome editing, and genome analysis, including transcriptome analysis, to analyze chrysanthemum, as well as the current status of and future prospects for chrysanthemum.

4.
Int J Mol Sci ; 24(4)2023 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-36835053

RESUMO

Sugarcane smut caused by Sporisorium scitamineum is one of the most devastating sugarcane diseases. Furthermore, Rhizoctonia solani causes severe diseases in various crops including rice, tomato, potato, sugar beet, tobacco, and torenia. However, effective disease-resistant genes against these pathogens have not been identified in target crops. Therefore, the transgenic approach can be used since conventional cross-breeding is not applicable. Herein, the overexpression of BROAD-SPECTRUM RESISTANCE 1 (BSR1), a rice receptor-like cytoplasmic kinase, was conducted in sugarcane, tomato and torenia. BSR1-overexpressing tomatoes exhibited resistance to the bacteria Pseudomonas syringae pv. tomato DC3000 and the fungus R. solani, whereas BSR1-overexpressing torenia showed resistance to R. solani in the growth room. Additionally, BSR1 overexpression conferred resistance to sugarcane smut in the greenhouse. These three BSR1-overexpressing crops exhibited normal growth and morphologies except in the case of exceedingly high levels of overexpression. These results indicate that BSR1 overexpression is a simple and effective tool for conferring broad-spectrum disease resistance to many crops.


Assuntos
Infecções Bacterianas , Oryza , Saccharum , Solanum lycopersicum , Ustilaginales , Oryza/genética , Saccharum/genética , Melhoramento Vegetal , Resistência à Doença/genética , Doenças das Plantas/microbiologia , Regulação da Expressão Gênica de Plantas
5.
Plant Biotechnol (Tokyo) ; 40(2): 157-165, 2023 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38250294

RESUMO

Chrysanthemum morifolium is one of the most popular ornamental plants in the world. However, as C. morifolium is a segmental hexaploid, self-incompatible, and has a sizable heterologous genome, it is difficult to modify its trait systematically. Genome editing technology is one of the attractive methods for modifying traits systematically. For the commercial use of genetically modified C. morifolium, rigorous stabilization of its quality is essential. This trait stability can be achieved by avoiding further genome modification after suitable trait modification by genome editing. Since C. morifolium is a vegetatively propagated plant, an approach for removing genome editing tools is required. In this study, we attempted to use the piggyBac transposon system to remove specific DNA sequences from the C. morifolium genome. Using the luminescence as a visible marker, we demonstrated that inoculation of Agrobacterium harboring hyperactive piggyBac transposase removes inserted 2.6 kb DNA, which harbors piggyBac recognition sequences, from the modified Eluc sequence.

6.
Sci Rep ; 12(1): 17874, 2022 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-36284128

RESUMO

Chrysanthemum morifolium is one of the most popular ornamental plants globally. Owing to its large and complex genome (around 10 Gb, segmental hexaploid), it has been difficult to obtain comprehensive transcriptome, which will promote to perform new breeding technique, such as genome editing, in C. morifolium. In this study, we used single-molecule real-time (SMRT) sequencing and RNA-seq technologies, combined them with an error-correcting process, and obtained high-coverage ray-floret transcriptome. The SMRT-seq data increased the ratio of long mRNAs containing complete open-reading frames, and the combined dataset provided a more complete transcriptomic data than those produced from either SMRT-seq or RNA-seq-derived transcripts. We finally obtained 'Sei Arabella' transcripts containing 928,645 non-redundant mRNA, which showed 96.6% Benchmarking Universal Single-Copy Orthologs (BUSCO) score. We also validated the reliability of the dataset by analyzing a mapping rate, annotation and transcript expression. Using the dataset, we searched anthocyanin biosynthesis gene orthologs and performed a qRT-PCR experiment to assess the usability of the dataset. The assessment of the dataset and the following analysis indicated that our dataset is reliable and useful for molecular biology. The combination of sequencing methods provided genetic information and a way to analyze the complicated C. morifolium transcriptome.


Assuntos
Chrysanthemum , Chrysanthemum/genética , Chrysanthemum/metabolismo , Transcriptoma , Regulação da Expressão Gênica de Plantas , Antocianinas/metabolismo , Flores/genética , Reprodutibilidade dos Testes , Perfilação da Expressão Gênica/métodos , Melhoramento Vegetal , Anotação de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala
7.
Int J Mol Sci ; 23(9)2022 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-35563126

RESUMO

Plant pathogens evade basal defense systems and attack different organs and tissues of plants. Genetic engineering of plants with genes that confer resistance against pathogens is very effective in pathogen control. Conventional breeding for disease resistance in ornamental crops is difficult and lagging relative to that in non-ornamental crops due to an inadequate number of disease-resistant genes. Therefore, genetic engineering of these plants with defense-conferring genes is a practical approach. We used rice BSR2 encoding CYP78A15 for developing transgenic Torenia fournieri Lind. lines. The overexpression of BSR2 conferred resistance against two devastating fungal pathogens, Rhizoctonia solani and Botrytis cinerea. In addition, BSR2 overexpression resulted in enlarged flowers with enlarged floral organs. Histological observation of the petal cells suggested that the enlargement in the floral organs could be due to the elongation and expansion of the cells. Therefore, the overexpression of BSR2 confers broad-spectrum disease resistance and induces the production of enlarged flowers simultaneously. Therefore, this could be an effective strategy for developing ornamental crops that are disease-resistant and economically more valuable.


Assuntos
Lamiales , Oryza , Resistência à Doença/genética , Flores/genética , Regulação da Expressão Gênica de Plantas , Lamiales/genética , Oryza/genética , Melhoramento Vegetal , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Plantas Geneticamente Modificadas/genética
8.
Planta ; 255(5): 105, 2022 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-35429252

RESUMO

MAIN CONCLUSION: A novel Torenia phenotype having separate petals was obtained by the combination of NF-YA6-VP16 with a floral organ-specific promoter. Genetic engineering techniques helped in obtaining novel flower colors and shapes, in particular, by introducing functionally modified transcription factors (TFs) to ornamental flower species. Herein, we used functionally modified Arabidopsis TFs fused with the repression domain SRDX and the activation domain VP16 to screen for novel floral traits in Torenia fournieri Lind (torenia). We avoided undesired phenotypes unrelated to flowers by expressing these TFs through a floral organ-specific promoter belonging to the class-B genes, GLOBOSA (TfGLO). Fourteen constructs were produced to express functionally modified Arabidopsis TFs in which each of SRDX and VP16 was fused into 7 TFs that were used for the collective transformation of Torenia plants. Among the obtained transgenic plants, phenotypes with novel floral traits reflected in separate petals within normally gamopetalous flower lines. Sequencing analysis revealed that the transgenic plants contained nuclear factor-YA6 (NF-YA6) fused with the VP16. In the margin between the lips of the petals and tube in the TfGLOp:NF-YA6-VP16 plants, staminoid organs have been developed to separate petals. In the petals of the TfGLOp:NF-YA6-VP16 plants, the expression of a Torenia class C gene, PLENA (TfPLE), was found to be ectopically increased. Moreover, expression of TfPLE-VP16 under the control of the TfGLO promoter brought a similar staminoid phenotype observed in the TfGLOp:NF-YA6-VP16 plants. These results suggest that the introduction of the TfGLOp:NF-YA6-VP16 induced TfPLE expression, resulting in the formation of staminoid petals and separation of them.


Assuntos
Arabidopsis , Lamiales , Arabidopsis/genética , Arabidopsis/metabolismo , Expressão Ectópica do Gene , Etoposídeo , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Lamiales/genética , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
9.
Planta ; 253(5): 100, 2021 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-33847818

RESUMO

MAIN CONCLUSION: Functional suppression of two types of class-C genes caused transformation of pistils and stamens into petaloid organs that exhibit novel phenotypes, which gives a distinct gorgeous impression in the florets of chrysanthemum. The multiple-petal trait is a breeding objective for many horticultural plants. The loss of function of class-C genes causes the multiple-petal trait in several plant species. However, mechanisms involved in the generation of the multiple-petal trait are unknown in Chrysanthemum morifolium (chrysanthemum). Here, we isolated 14 class-C AGAMOUS (AG) genes, which were classified into two types of class-C genes, in chrysanthemum. Seven of these were categorized into CAG type 1 genes (CAG1s) and seven into CAG type 2 genes (CAG2s). Functions of class-C genes were co-suppressed by chimeric repressors and simultaneously knocked-down by RNAi to produce the multiple-petal phenotype in chrysanthemum. The expression of chimeric repressors of CAG1s and CAG2s caused morphological alteration of the pistils and stamens into petaloid organs in the ray and disk florets. Interestingly, the reproductive organs of the disk florets were transformed into petaloid organs similar to the petals of the disk florets, and those of the ray florets were transformed into petaloid organs such as the petals of the ray florets. Simultaneous knockdown of CAG1s and CAG2s expression by RNAi also exhibited a petaloid phenotype as observed in transgenic plants obtained by chimeric repressors. These results showed that CAG1s and CAG2s play important roles in the development of pistils and stamens, and the simultaneous repression of CAG1s and CAG2s resulted in a multiple-petal phenotype in chrysanthemum.


Assuntos
Chrysanthemum , Chrysanthemum/genética , Chrysanthemum/metabolismo , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas , Genitália/metabolismo , Fenótipo , Melhoramento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Interferência de RNA
10.
Plant Biotechnol (Tokyo) ; 37(3): 373-375, 2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-33088204

RESUMO

The shoot apical meristem (SAM) is typically divided into three cell layers: the outermost epidermal layer (L1), the subepidermal layer (L2) and the inner corpus region (L3). Structures within the cell layers are normally maintained throughout development; however, through vegetative propagation of a periclinal chimeric chrysanthemum expressing a fluorescent protein gene only in the L1 layer, we collected twelve independent shoots that had partially mosaic fluorescent inner cells (L2, L3) in addition to fluorescent epidermal cells (L1). Furthermore, the elongated tissues of nine shoots out of the twelve had no internal fluorescent cells, i.e., they had the original L1 chimerism. Observations of the fluorescence distribution suggested that the change in chimerism occurred at the nodes, indicating previously unnoticed cell layer dynamics occurring at the nodes.

11.
PLoS One ; 15(8): e0237173, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32845897

RESUMO

Gentian is an important ornamental flower in Japan. The corolla of the majority of cultivated Japanese gentians have green spots, which are rarely encountered in flowers of other angiosperms. Little information is available on the functional traits of the green spots. In this study, we characterized the green spots in the Japanese gentian corolla using a number of microscopic techniques. Opto-digital microscopy revealed that a single visible green spot is composed of approximately 100 epidermal cells. The epidermal cells of a green spot formed a dome-like structure and the cell lumen contained many green structures that were granular and approximately 5 µm in diameter. The green structures emitted red autofluorescence when irradiated with 488 nm excitation light. Transmission electron microscopy revealed that the green structures contained typical thylakoids and grana, thus indicating they are chloroplasts. No grana were observed and the thylakoids had collapsed in the plastids of epidermal cells surrounding green spots. To estimate the rate of photosynthetic electron transfer of the green spots, we measured chlorophyll fluorescence using the MICROSCOPY version of an Imaging-PAM (pulse-amplitude-modulated) fluorometer. Under actinic light of 449 µmol m-2 s-1, substantial electron flow through photosystem II was observed. Observation of green spot formation during corolla development revealed that immature green spots formed at an early bud stage and developed to maturity associated with chloroplast degradation in the surrounding epidermal cells. These results confirmed that the Japanese gentian corolla contains functional chloroplasts in restricted areas of epidermal cells and indicated that a sophisticated program for differential regulation of chloroplast formation and degradation is operative in the epidermis.


Assuntos
Flores/citologia , Flores/metabolismo , Gentiana/anatomia & histologia , Tilacoides/metabolismo , Clorofila/metabolismo , Transporte de Elétrons , Japão , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Fotossíntese , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Epiderme Vegetal/citologia , Epiderme Vegetal/metabolismo , Folhas de Planta/metabolismo
12.
Planta ; 251(5): 101, 2020 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-32333191

RESUMO

MAIN CONCLUSION: Simultaneous knockdown or knockout of Torenia fournieri PLENA (TfPLE) and FALINELLI (TfFAR) genes with RNAi or genome-editing technologies generated a multi-petal phenotype in torenia. The MADS-box gene AGAMOUS (AG) is well known to play important roles in the development of stamens and carpels in Arabidopsis. Mutations in AG cause the morphological transformation of stamens and carpels into petaloid organs. In contrast, torenia (Torenia fournieri Lind.) has two types of class-C MADS-box genes, PLENA (PLE) and FALINELLI (FAR); however, their functions were previously undetermined. To examine the function of TfPLE and TfFAR in torenia, we used RNAi to knockdown expression of these two genes. TfPLE and TfFAR double-knockdown transgenic torenia plants had morphologically altered stamens and carpels that developed into petaloid organs. TfPLE knockdown transgenic plants also exhibited morphological transformations that included shortened styles, enlarged ovaries, and absent stigmata. Furthermore, simultaneous disruption of TfPLE and TfFAR genes by CRISPR/Cas9-mediated genome editing also resulted in the conversion of stamens and carpels into petaloid organs as was observed in the double-knockdown transgenic plants mediated by RNAi. In addition, the carpels of one TfPLE knockout mutant had the same morphological abnormalities as TfPLE knockdown transgenic plants. TfFAR knockdown genome-edited mutants had no morphological changes in their floral organs. These results clearly show that TfPLE and TfFAR cooperatively play important roles in the development of stamens and carpels. Simultaneous disruption of TfPLE and TfFAR functions caused a multi-petal phenotype, which is expected to be a highly valuable commercial floral trait in horticultural flowers.


Assuntos
Arabidopsis/genética , Edição de Genes , Lamiales/genética , Proteínas de Domínio MADS/genética , Interferência de RNA , Arabidopsis/crescimento & desenvolvimento , Flores/genética , Flores/crescimento & desenvolvimento , Técnicas de Silenciamento de Genes , Lamiales/crescimento & desenvolvimento , Proteínas de Domínio MADS/metabolismo , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas
13.
Plant Cell Rep ; 39(5): 609-619, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32060603

RESUMO

KEY MESSAGE: A fluorescent protein visualized distributions of cell layers in floral organs of chrysanthemum using transgenic periclinal chimeras carrying a gene encoding a fluorescent compound. Plant meristems have three cell layers: the outermost layer (L1), the second layer (L2), and the inner layer (L3). The layers are maintained during development but there is limited knowledge of the details of cell layer patterns within floral organs. In this study, we visualized the distributions of cell layers in floral organs of chrysanthemum using periclinal chimeras carrying a gene encoding a fluorescent compound in the L1 or the L2/L3 layers. The L1 layer contributed most of the epidermal cells of organs including the receptacle, petal, anther, filament, style, stigma, and ovule. The transmitting tissue in the pistil and most of the internal area of the ovule were also derived from the L1. In crossing experiments, no progeny of the L1-chimeric plants showed fluorescence, indicating that the germ cells of chrysanthemum are not derived from the L1 layer. Since anthocyanin pigment is present only in the L1-derived epidermal cells of petals, L1-specific gene integration could be used to alter flower color in commercial cultivars, with a reduced risk of transgene flow from the transgenic chrysanthemums to wild relatives.


Assuntos
Chrysanthemum/crescimento & desenvolvimento , Chrysanthemum/genética , Flores/citologia , Proteínas Luminescentes/genética , Meristema/citologia , Antocianinas/metabolismo , Quimera/genética , Quimera/metabolismo , Chrysanthemum/citologia , Troca Genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Células Germinativas Vegetais/citologia , Células Germinativas Vegetais/metabolismo , Proteínas Luminescentes/metabolismo , Meristema/crescimento & desenvolvimento , Pigmentação , Epiderme Vegetal/citologia , Plantas Geneticamente Modificadas , Transgenes
14.
Breed Sci ; 69(3): 536-544, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31598089

RESUMO

The chrysanthemum (Chrysanthemum morifolium) is one of the most popular ornamental plants in the world. Genetic transformation is a promising tool for improving traits, editing genomes, and studying plant physiology. Promoters are vital components for efficient transformation, determining the level, location, and timing of transgene expression. The cauliflower mosaic virus (CaMV) 35S promoter is most frequently used in dicotyledonous plants but is less efficient in chrysanthemums than in tobacco or torenia plants. Previously, we used the parsley ubiquitin (PcUbi) promoter in chrysanthemums for the first time and analyzed its activity in transgenic calli. To expand the variety of constitutive promoters in chrysanthemums, we cloned the upstream region of the actin 2 (CmACT2) gene and compared its promoter activity with the 35S and PcUbi promoters in several organs, as well as its durability for long-term cultivation. The CmACT2 promoter has higher activity than the 35S promoter in calli but is less durable. The PcUbi promoter has the highest activity not only in calli but also in leaves, ray florets, and disk florets, and retains its activity after long-term cultivation. In conclusion, we have provided useful information and an additional type of promoter available for transgene expression in chrysanthemums.

15.
Plant Physiol Biochem ; 131: 47-52, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29709514

RESUMO

Ornamental plants, like roses, carnations, and chrysanthemums, are economically important and are sold all over the world. In addition, numerous cut and garden flowers add colors to homes and gardens. Various strategies of plant breeding have been employed to improve traits of many ornamental plants. These approaches span from conventional techniques, such as crossbreeding and mutation breeding, to genetically modified plants. Recently, genome editing has become available as an efficient means for modifying traits in plant species. Genome editing technology is useful for genetic analysis and is poised to become a common breeding method for ornamental plants. In this review, we summarize the benefits and limitations of conventional breeding techniques and genome editing methods and discuss their future potential to accelerate the rate breeding programs in ornamental plants.


Assuntos
Engenharia Genética/métodos , Plantas/genética , Sistemas CRISPR-Cas/genética , Previsões , Engenharia Genética/tendências , Genoma de Planta/genética , Melhoramento Vegetal/métodos , Plantas Geneticamente Modificadas/genética
16.
Breed Sci ; 68(1): 88-98, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29681751

RESUMO

Transcription factors play important roles not only in the development of floral organs but also in the formation of floral characteristics in various plant species. Therefore, transcription factors are reasonable targets for modifying these floral traits and generating new flower cultivars. However, it has been difficult to control the functions of transcription factors because most plant genes, including those encoding transcription factors, exhibit redundancy. In particular, it has been difficult to understand the functions of these redundant genes by genetic analysis. Thus, a breakthrough silencing method called chimeric repressor gene silencing technology (CRES-T) was developed specifically for plant transcription factors. This method transforms transcriptional activators into dominant repressors, and the artificial chimeric repressors suppress the function of transcription factors regardless of their redundancy. Among these chimeric repressors, some were found to be inappropriate for expression throughout the plant body because they resulted in deformities. For these chimeric repressors, utilization of floral organ-specific promoters overcomes this problem by avoiding expression throughout the plant body. In contrast, attachment of viral activation domain VP16 to transcriptional repressors effectively alters into transcriptional activators. This review presents the importance of transcription factors for characterizing floral traits, describes techniques for controlling the functions of transcription factors.

18.
BMC Plant Biol ; 17(1): 202, 2017 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-29141585

RESUMO

BACKGROUND: Chlorophylls (Chls) are magnesium-containing tetrapyrrole macromolecules responsible for the green color in plants. The Chl metabolic pathway has been intensively studied and nearly all the enzymes involved in the pathway have been identified and characterized. Synthesis and activity of these enzymes are tightly regulated in tissue- and developmental stage-specific manners. Leaves contain substantial amounts of Chls because Chls are indispensable for photosynthesis. In contrast, petals generally contain only trace amounts of Chls, which if present would mask the bright petal color. Limited information is available about the mechanisms that control such tissue-specific accumulation of Chls. RESULTS: To identify the regulatory steps that control Chl accumulation, we compared gene expression in petals and leaves of chrysanthemum cultivars with different Chl levels. Microarray and quantitative real-time PCR analyses showed that the expression levels of Chl biosynthesis genes encoding glutamyl-tRNA reductase, Mg-protoporphyrin IX chelatase, Mg-protoporphyrin IX monomethylester cyclase, and protochlorophyllide oxidoreductase were well associated with Chl content: their expression levels were lower in white petals than in green petals, and were highest in leaves. Among Chl catabolic genes, expression of STAY-GREEN, encoding Mg-dechelatase, which is a key enzyme controlling Chl degradation, was considerably higher in white and green petals than in leaves. We searched for transcription factor genes whose expression was well related to Chl level in petals and leaves and found three such genes encoding MYB113, CONSTANS-like 16, and DREB and EAR motif protein. CONCLUSIONS: From our transcriptome analysis, we assume that a low rate of Chl biosynthesis and a high rate of Chl degradation lead to the absence of Chls in white chrysanthemum petals. We identified several candidate transcription factors that might affect Chl accumulation in chrysanthemum petals. Functional analysis of these transcription factors will provide a basis for future molecular studies of tissue-specific Chl accumulation.


Assuntos
Clorofila/metabolismo , Chrysanthemum/metabolismo , Flores/metabolismo , Folhas de Planta/metabolismo , Clorofila/análise , Chrysanthemum/genética , Flores/química , Perfilação da Expressão Gênica , Genes de Plantas , Redes e Vias Metabólicas , Microscopia Eletrônica de Transmissão , Análise de Sequência com Séries de Oligonucleotídeos , Folhas de Planta/química , Reação em Cadeia da Polimerase em Tempo Real
19.
BMC Genomics ; 18(1): 683, 2017 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-28870156

RESUMO

BACKGROUND: Chrysanthemum morifolium is one of the most economically valuable ornamental plants worldwide. Chrysanthemum is an allohexaploid plant with a large genome that is commercially propagated by vegetative reproduction. New cultivars with different floral traits, such as color, morphology, and scent, have been generated mainly by classical cross-breeding and mutation breeding. However, only limited genetic resources and their genome information are available for the generation of new floral traits. RESULTS: To obtain useful information about molecular bases for floral traits of chrysanthemums, we read expressed sequence tags (ESTs) of chrysanthemums by high-throughput sequencing using the 454 pyrosequencing technology. We constructed normalized cDNA libraries, consisting of full-length, 3'-UTR, and 5'-UTR cDNAs derived from various tissues of chrysanthemums. These libraries produced a total number of 3,772,677 high-quality reads, which were assembled into 213,204 contigs. By comparing the data obtained with those of full genome-sequenced species, we confirmed that our chrysanthemum contig set contained the majority of all expressed genes, which was sufficient for further molecular analysis in chrysanthemums. CONCLUSION: We confirmed that our chrysanthemum EST set (contigs) contained a number of contigs that encoded transcription factors and enzymes involved in pigment and aroma compound metabolism that was comparable to that of other species. This information can serve as an informative resource for identifying genes involved in various biological processes in chrysanthemums. Moreover, the findings of our study will contribute to a better understanding of the floral characteristics of chrysanthemums including the myriad cultivars at the molecular level.


Assuntos
Chrysanthemum/anatomia & histologia , Chrysanthemum/genética , Etiquetas de Sequências Expressas/metabolismo , Flores/anatomia & histologia , Genes de Plantas/genética , Sequenciamento de Nucleotídeos em Larga Escala , Sequência de Aminoácidos , Carotenoides/metabolismo , Anotação de Sequência Molecular , Terpenos/metabolismo , Fatores de Transcrição/genética
20.
Hortic Res ; 4: 17008, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28446955

RESUMO

Although chimeric repressors such as the Arabidopsis TCP3 repressor are known to have significant effects on flower morphology and color, their cellular-level effects on flower petals are not understood. The promoter sequences of the genes expressed in the flowers of cyclamen, a representative potted flower grown during the winter season, are also unknown. Here, we isolated eight promoters from cyclamen genes that are reportedly expressed in the petals. These promoters were then fused to four chimeric repressors and introduced into the model flower torenia to screen for effective combinations of promoters and repressors for flower breeding. As expected, some of the constructs altered flower phenotypes upon transformation. We further analyzed the effects of chimeric repressors at the cellular level. We observed that complicated petal and leaf serrations were accompanied by excessive vascular branching. Dichromatism in purple anthocyanin was inferred to result in bluish flowers, and imbalanced cell proliferation appeared to result in epinastic flowers. Thus, the genetic constructs and phenotypic changes described in this report will benefit the future breeding and characterization of ornamental flowers.

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