Characterization of Three Chalcone Synthase-like Genes in Dianthus Chinensis

Abstract Being an important garden plant, Dianthus chinensis flower has a great variety of colors and color patterns. Chalcone synthase (CHS) is the key enzyme in the anthocyanin biosynthetic pathway. Although CHS genes have been isolated and characterized in ornamental plants, the CHS gene is still unknown in D. chinensis. In our study, three CHS genes, DchCHS1 (KX893854), DchCHS2 (MK404175) and DchCHS3 (MK416198) were isolated in D. chinensis. Their deduced amino acid sequences show high homology with the known CHS sequences in Caryophyllaceae. The phylogenetic tree suggests that the DchCHS1 and the DchCHS3 have a close relation with the known CHS sequences in Caryophyllaceae and the DchCHS2 is different from them. The DchCHSs were characterized by the Tobacco Rattle Virus (TRV)-based virus-induced gene silencing (VIGS) system. We obtained white or pale purple flowers in the DchCHS1-silenced flowers and reducing purple flowers in the DchCHS2-silenced and the DchCHS3-silenced flowers. The anthocyanin content and the transcript level of the silenced DchCHS were significantly reduced in accordance with the silencing phenotypes. The DchCHSs showed different expression patterns during floral bud developments, among flower colors and in organs. Their expression levels in the purple flower were greatly higher than those in the white flower. Compared with DchCHS2 and DchCHS3, DchCHS1 was abundantly expressed at each floral bud stage, in each flower color and in the flower organ. In conclusion, the three DchCHSs are all involved in the anthocyanin synthesis and the flower coloration, and DchCHS1 probably plays a major role in D. chinensis flowers.

Download Full-text

DFR and PAL gene transcription and their correlation with anthocyanin accumulation in Rhodomyrtus tomentosa (Aiton.) Hassk.

Turkish Journal of Biochemistry ◽

10.1515/tjb-2018-0019 ◽

2018 ◽

Vol 44 (3) ◽

pp. 289-298

Author(s):

Bao-Jun Zhu ◽

Qian Wang ◽

Jing-Hui Wang ◽

Lin-Lin Gao ◽

Jing-Wen Zhang ◽

...

Keyword(s):

Expression Patterns ◽

Pcr Analysis ◽

Anthocyanin Synthesis ◽

Anthocyanin Content ◽

Transcript Levels ◽

Color Development ◽

Phenylalanine Ammonialyase ◽

Maturity Period ◽

Late Maturity ◽

Rhodomyrtus Tomentosa

Abstract Objectives Rhodomyrtus tomentosa (Aiton.) Hassk. (R. tomentosa) is rich in nutrients and has multiple pharmacological applications. Anthocyanins confer color to the flowers and berries of R. tomentosa and provide protection against photodamage. The dihydroflavonol 4-reductase gene (DFR) and phenylalanine ammonialyase gene (PAL) are crucial for anthocyanin synthesis. Methods DFR and PAL transcript levels and anthocyanin content in the pigmented organs of R. tomentosa were investigated through qRT-PCR analysis and spectrophotometry, respectively. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was selected as the reference gene for the normalization of DFR and PAL transcript levels. Results Transcript levels of DFR and PAL were higher in organs with vigorous metabolism than those in senescent organs. DFR and PAL transcript levels were up-regulated during the initial and middle-maturity periods of fruit. These expression patterns are consistent with fruit color development. The highest transcript levels of PAL and DFR were observed during the middle-maturity period or the red-fruit period. Conclusion During the late maturity period of R. tomentosa fruit, the transcript levels of the two genes were down-regulated even though anthocyanins were continuously accumulated, which was different from the accumulation of anthocyanins in some late mature fruits.

Download Full-text

Expression characterisation of cyclophilin BrROC1 during light treatment and abiotic stresses response in Brassica rapa subsp. rapa ‘Tsuda’

Functional Plant Biology ◽

10.1071/fp18029 ◽

2018 ◽

Vol 45 (12) ◽

pp. 1223 ◽

Cited By ~ 3

Author(s):

Haifang Yan ◽

Bo Zhou ◽

Wei He ◽

Yuzhe Nie ◽

Yuhua Li

Keyword(s):

Blue Light ◽

Brassica Rapa ◽

Abiotic Stresses ◽

Expression Patterns ◽

Transcript Level ◽

Light Treatment ◽

Anthocyanin Synthesis ◽

Root Epidermis ◽

High Or Low Temperature ◽

Level Analysis

ROC1 is a prototypic peptidyl prolyl cis/trans isomerase (PPIase) of the plant cytosol belonging to the large subfamily of cyclophilins that are associated with diverse functions through foldase, scaffolding, chaperoning or other unknown activities. Although many functions of plant cyclophilins have been reported, the molecular basis of stress-responsive expression of plant cyclophilins is still largely unknown. To characterise the roles of BrROC1 during light treatment and their responses in various abiotic stresses, we identified BrROC1 genes and characterised their expression patterns in Brassica rapa subsp. rapa ‘Tsuda’. Our results showed that BrROC1 genes are multi-family genes. Transcript level analysis showed BrROC1-2 expressed higher than BrROC1-1 in 0 to 6-day-old seedlings under natural light. Moreover, BrROC1-2 genes were also induced to highly express in the cotyledon, upper hypocotyls and lower hypocotyls of seedlings under UV-A and blue-light treatment. In addition, the transcript level of BrROC1-1 was higher in pigment tissues than that in unpigment tissues (cotyledon and lower hypocotyl) under UV-A and blue-light treatment. Furthermore, when the unpigment epidermis (shaded light) of 2-month-old ‘Tsuda’ turnip roots was exposed to UV-A light, transcript levels of the BrROC1-1 and BrROC1-2 were significantly increased with time prolongation. These two BrROC1 genes might be involved in UV-A-induced anthocyanin synthesis in the root epidermis of ‘Tsuda’ turnip, which accumulates high levels of anthocyanin. These two BrROC1 genes were also induced to be regulated by abiotic stresses such as high or low temperature, dehydration, osmotic and salt stresses. Then, the results indicate that BrROC1 genes are involved in light induction response and may play important roles in adaptation of plants to various environmental stresses.

Download Full-text

Transcriptome Analysis Revealed the Mechanism by Which Exogenous ABA Increases Anthocyanins in Blueberry Fruit During Veraison

Frontiers in Plant Science ◽

10.3389/fpls.2021.758215 ◽

2021 ◽

Vol 12 ◽

Author(s):

Tianyu Han ◽

Wenlong Wu ◽

Weilin Li

Keyword(s):

Transcription Factors ◽

Fruit Ripening ◽

Expression Patterns ◽

Synthesis Process ◽

Mature Stage ◽

Anthocyanin Synthesis ◽

Bhlh Transcription Factor ◽

Anthocyanin Content ◽

Anthocyanin Pathway ◽

Green Stage

Blueberry (Vaccinium spp.) is a popular healthy fruit worldwide. The health value of blueberry is mainly because the fruit is rich in anthocyanins, which have a strong antioxidant capacity. However, because blueberry is a non-model plant, little is known about the structural and regulatory genes involved in anthocyanin synthesis in blueberries. Previous studies have found that spraying 1,000 mg/L abscisic acid at the late green stage of “Jersey” highbush blueberry fruits can increase the content of anthocyanins. In this experiment, the previous results were verified in “Brightwell” rabbiteye blueberry fruits. Based on the previous results, the anthocyanin accumulation process in blueberry can be divided into six stages from the late green stage to the mature stage, and the transcriptome was used to systematically analyze the blueberry anthocyanin synthesis process. Combined with data from previous studies on important transcription factors regulating anthocyanin synthesis in plants, phylogenetic trees were constructed to explore the key transcription factors during blueberry fruit ripening. The results showed that ABA increased the anthocyanin content of blueberry fruits during veraison. All structural genes and transcription factors (MYB, bHLH, and WD40) involved in the anthocyanin pathway were identified, and their spatiotemporal expression patterns were analyzed. The expression of CHS, CHI, DFR, and LDOX/ANS in ABA-treated fruits was higher in the last two stages of maturity, which was consistent with the change in the anthocyanin contents in fruits. In general, six MYB transcription factors, one bHLH transcription factor and four WD40 transcription factors were found to change significantly under treatment during fruit ripening. Among them, VcMYBA plays a major role in the regulation of anthocyanin synthesis in ABA signaling. This result preliminarily explained the mechanism by which ABA increases the anthocyanin content and improves the efficiency of the industrial use of blueberry anthocyanins.

Download Full-text

Chalcone Synthase-Encoding AeCHS is Involved in Normal Petal Coloration in Actinidia eriantha

Genes ◽

10.3390/genes10120949 ◽

2019 ◽

Vol 10 (12) ◽

pp. 949

Author(s):

Yukuo Li ◽

Wen Cui ◽

Xiujuan Qi ◽

Chengkui Qiao ◽

Miaomiao Lin ◽

...

Keyword(s):

Gene Expression ◽

Structural Gene ◽

Gene Expression Analysis ◽

Chalcone Synthase ◽

Anthocyanin Biosynthesis ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Anthocyanin Content ◽

Virus Induced Gene Silencing

Studies on anthocyanin biosynthesis have been mainly concentrated on the fruit, whereas few have focused the mechanism of flower coloration in kiwifruit. Here, we report that the structural gene, AeCHS, is involved in anthocyanin accumulation and indispensable for normal petal coloration in Actinidia eriantha. Petals from three different species including Actinidia eriantha (red petals), Actinidia hemsleyana (light pink petals) and Actinidia arguta (white petals) were selected for anthocyanin determination and gene expression analysis. The anthocyanin components in A. eriantha were significantly higher than in A. hemsleyana or A. arguta. Consistently, gene expression profiles suggested that AeCHS expression in A. eriantha was higher than in A. hemsleyana or A. arguta. Cluster analysis showed that AeCHS was clustered into a single group and distinctly separated from other genes, indicating the expression pattern of AeCHS gene was different from any other. Additionally, correlation analysis revealed AeCHS expression significantly correlated with anthocyanin content. The complete coding sequence of AeCHS was cloned from petals of A. eriantha ‘Zaoxu’, showing the length of AeCHS was 1170 bp encoding a protein of 389 amino acids. AeCHS was located in the cytoplasm, indicating it is indeed a structural gene involved in anthocyanin biosynthesis. AeCHS silencing performed by infiltration grafting-mediated virus-induced gene silencing (VIGS) reduced petal anthocyanin content and bleached red petals in A. eriantha. Our results confirm a crucial role of AeCHS in anthocyanin biosynthesis and accumulation in A. eriantha petals; furthermore, they offer important basic information and constitute a reference point for further research.

Download Full-text

Transcriptome Co-Expression Network Analysis Identifies Key Genes and Regulators of Sweet Cherry Anthocyanin Biosynthesis

Horticulturae ◽

10.3390/horticulturae7060123 ◽

2021 ◽

Vol 7 (6) ◽

pp. 123

Author(s):

Haiying Yang ◽

Changping Tian ◽

Xiwen Li ◽

Hansheng Gong ◽

Aidi Zhang

Keyword(s):

Transcription Factors ◽

Network Analysis ◽

Sweet Cherry ◽

Anthocyanin Biosynthesis ◽

Expression Patterns ◽

Luciferase Assay ◽

Anthocyanin Synthesis ◽

Anthocyanin Content ◽

Sweet Cherries ◽

Key Factor

Anthocyanin is the key factor that results in the attractive color of sweet cherry fruits. However, information regarding sweet cherry coloration and the potential mechanisms underlying anthocyanin biosynthesis is limited. In this study, we found that the anthocyanin accumulation varied in sweet cherry flesh and peel, while the anthocyanin content increased sharply in the dark red (DR) stage. Correlations between anthocyanin concentrations and RNA sequencing (RNA-seq), constructed with Weighted Gene Co-Expression Network Analysis (WGCNA), indicated that two structural genes (Pac4CL2, PacANS) and 11 transcription factors (PacbHLH13/74, PacDIV, PacERF109/115, PacGATA8, PacGT2, PacGTE10, PacMYB308, PacPosF21, and PacWRKY7) had similar expression patterns with the changes in anthocyanin content. Additionally, real-time PCR verified all of these gene expression patterns and revealed that PacANS exhibited the highest transcription level. In order to search for potential regulators for anthocyanin biosynthesis, a dual-luciferase assay was performed to investigate the regulatory activities of 11 transcription factors on the PacANS promoter. The results revealed that two novelty bHLHs, PacbHLH13 and PacbHLH74, can trans-activate the PacANS promoter and they might be the candidate genes for regulating anthocyanin synthesis in sweet cherry fruits. The present findings provide a novel viewpoint with regard to anthocyanin biosynthesis mechanisms and the regulatory transcriptional network of fruit coloration in sweet cherries.

Download Full-text

Genome-wide analysis of chalcone synthase (CHS) family from eggplant (Solanum melongena L.) in flavonoid biosynthetic pathway and expression pattern in response to heat stress

10.21203/rs.2.17462/v1 ◽

2019 ◽

Author(s):

Xuexia Wu ◽

Dingshi Zha

Keyword(s):

Heat Stress ◽

Chalcone Synthase ◽

Expression Patterns ◽

Regulatory Elements ◽

Regulation Mechanism ◽

Anthocyanin Content ◽

Genome Wide ◽

Peel Color ◽

Solanaceae Species ◽

Key Genes

Abstract Background Enzymes of chalcone synthase (CHS) family participate in the synthesis of a series of secondary metabolites in plants, fungi and bacteria. Some of these metabolites have a wide variety of biological functions such as flower pigmentation, protection against UV radiation, pathogen defense, auxin transport and pollen fertility. CHS also showed significant correlation to the accumulation patterns of anthocyanin. The peel color mainly determined by the content of anthocyanin, is a majority economic trait for eggplant affected by heat stress. Results A total of 7 CHS ( SmCHS1-7 ) putative genes were identified in genome-wide of eggplant ( S. melongena L . ). The SmCHS genes distribute on 7 scaffolds and were classified into 3 clusters. Phylogenetic relationships analysis showed that 73 CHS genes from 7 Solanaceae species were classified into 10 groups. SmCHS5 , SmCHS6 and SmCHS7 were continuously down-regulated under 38℃ and 45℃ treatment, while SmCHS4 was up-regulated under 38℃ but little change at 45℃ in peel. Expression profiles of anthocyanin biosynthesis key genes families showed that the PAL, 4CL and AN11 genes were mainly expressed in all five tissues. CHI, F3H, F3’5’H, DFR, 3GT and bHLH1 genes were expressed in flower and peel. Under heat stress, 52 key genes expression level were reduced under heat stress. By contrast, expression patterns of eight key genes similar to SmCHS4 up-regulated at the 38℃-3h. Conclusions Comparative analysis of putative CHS protein biochemical characteristics, cis -regulatory elements, regulatory network revealed that SmCHS genes family have conservation gene structure and functional diversification. SmCHS showed two or more expression patterns and execute multiple functions to regulate anthocyanin content. Combined with regulatory networks, it is possible to further understand the regulation mechanism of peel color in eggplant.

Download Full-text

Transcriptome Analysis Revealed The Mechanism of Exogenous ABA Increasing Anthocyanins in Blueberry Fruit During Veraison

10.21203/rs.3.rs-88852/v1 ◽

2020 ◽

Author(s):

Tianyu Han ◽

Zhixiang Yan ◽

Wenlong Wu ◽

Weilin Li

Keyword(s):

Transcription Factors ◽

Transcriptome Analysis ◽

Expression Patterns ◽

Synthesis Process ◽

Anthocyanin Synthesis ◽

Bhlh Transcription Factor ◽

Anthocyanin Content ◽

Structural Genes ◽

Anthocyanin Pathway ◽

Green Stage

Abstract Background: Blueberry（Vaccinium Spp）is a popular healthy fruit all over the world. The health value of blueberry is mainly due to the fact that blueberry is rich in anthocyanins, which have a strong antioxidant capacity. However, due to the fact that blueberry is a non model plant, little is known about the structural genes and regulatory genes involved in the anthocyanin synthesis of blueberries. Previous studies have found that spraying abscisic acid at the late green stage of blueberry fruit can increase the content of anthocyanins. Based on the former results, the anthocyanin accumulation process of blueberry can be divided into six stages from late green stage to mature stage to analyze the anthocyanin synthesis mechanism. In order to identify the important genes in the anthocyanin synthesis process of blueberry, the transcriptome analysis was conducted to explore the key genes in blueberry anthocyanin synthesis process.Results: The results showed that ABA could increase the anthocyanin content of blueberry fruits during the veraison. The effect of ABA on blueberry fruit development was systematically analyzed by KEGG and GO. All structural genes and transcription factors (MYB, bHLH and WD40) involved in anthocyanin pathway were identified and their spatiotemporal expression patterns were analyzed. The expression of CHS, CHI, DFR and LDOX / ANS in ABA treated fruits was higher in S5-S6, which was consistent with the change of anthocyanins in fruits.In general, six MYB transcription factors, one bHLH transcription factor and four WD40 transcription factors under treatment were found to have significant changes in transcripts during fruit ripening. Conclusions: Our results suggest that VcMYBA should play a major role in the regulation of anthocyanin synthesis in ABA signaling. This result preliminarily explained the mechanism of ABA increasing anthocyanin content and improves the efficiency of industrial use of blueberry anthocyanins.

Download Full-text

Genome-wide analysis of chalcone synthase (CHS) family from eggplant (Solanum melongena L.) in flavonoid biosynthetic pathway and expression pattern in response to heat stress

10.1101/863696 ◽

2019 ◽

Author(s):

Xuexia Wu ◽

Shengmei Zhang ◽

Xiaohui Liu ◽

Jing Shang ◽

Aidong Zhang ◽

...

Keyword(s):

Heat Stress ◽

Chalcone Synthase ◽

Expression Patterns ◽

Regulatory Elements ◽

Regulation Mechanism ◽

Anthocyanin Content ◽

Genome Wide ◽

Peel Color ◽

Solanaceae Species ◽

Key Genes

AbstractEnzymes of chalcone synthase (CHS) family participate in the synthesis of a series of secondary metabolites in plants, fungi and bacteria. CHS showed significant correlation to the accumulation patterns of anthocyanin. The peel color mainly determined by the content of anthocyanin, is a majority economic trait for eggplant affected by heat stress. A total of 7 CHS(SmCHS1-7) putative genes were identified in genome-wide of eggplant (S. melongena L.). The SmCHS genes distribute on 7 scaffolds and were classified into 3 clusters. Phylogenetic relationships analysis showed that 73 CHS genes from 7 Solanaceae species were classified into 10 groups. SmCHS5, SmCHS6 and SmCHS7 were continuously down-regulated under 38°C and 45°C treatment, while SmCHS4 was up-regulated under 38°C but little change at 45°C in peel. Expression profiles of anthocyanin biosynthesis key genes families showed that the PAL, 4CL and AN11 genes were mainly expressed in all five tissues. CHI, F3H, F3’5’H, DFR, 3GT and bHLH1 genes were expressed in flower and peel. Under heat stress, 52 key genes expression level were reduced under heat stress. By contrast, expression patterns of eight key genes similar to SmCHS4 up-regulated at the 38°C-3h. Comparative analysis of putative CHS protein biochemical characteristics, cis-regulatory elements, regulatory network revealed that SmCHS genes family have conservation gene structure and functional diversification. SmCHS showed two or more expression patterns and execute multiple functions to regulate anthocyanin content. Combined with regulatory networks, it is possible to further understand the regulation mechanism of peel color in eggplant.

Download Full-text

Roles of the 14-3-3 gene family in cotton flowering

BMC Plant Biology ◽

10.1186/s12870-021-02923-9 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Na Sang ◽

Hui Liu ◽

Bin Ma ◽

Xianzhong Huang ◽

Lu Zhuo ◽

...

Keyword(s):

Gene Family ◽

Protein Interactions ◽

Leucine Zipper ◽

Expression Patterns ◽

Bimolecular Fluorescence Complementation ◽

Structural Motif ◽

Virus Induced Gene Silencing ◽

Protein Protein Interactions ◽

Basic Leucine Zipper ◽

Genome Wide

Abstract Background In plants, 14-3-3 proteins, also called GENERAL REGULATORY FACTORs (GRFs), encoded by a large multigene family, are involved in protein–protein interactions and play crucial roles in various physiological processes. No genome-wide analysis of the GRF gene family has been performed in cotton, and their functions in flowering are largely unknown. Results In this study, 17, 17, 31, and 17 GRF genes were identified in Gossypium herbaceum, G. arboreum, G. hirsutum, and G. raimondii, respectively, by genome-wide analyses and were designated as GheGRFs, GaGRFs, GhGRFs, and GrGRFs, respectively. A phylogenetic analysis revealed that these proteins were divided into ε and non-ε groups. Gene structural, motif composition, synteny, and duplicated gene analyses of the identified GRF genes provided insights into the evolution of this family in cotton. GhGRF genes exhibited diverse expression patterns in different tissues. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that the GhGRFs interacted with the cotton FLOWERING LOCUS T homologue GhFT in the cytoplasm and nucleus, while they interacted with the basic leucine zipper transcription factor GhFD only in the nucleus. Virus-induced gene silencing in G. hirsutum and transgenic studies in Arabidopsis demonstrated that GhGRF3/6/9/15 repressed flowering and that GhGRF14 promoted flowering. Conclusions Here, 82 GRF genes were identified in cotton, and their gene and protein features, classification, evolution, and expression patterns were comprehensively and systematically investigated. The GhGRF3/6/9/15 interacted with GhFT and GhFD to form florigen activation complexs that inhibited flowering. However, GhGRF14 interacted with GhFT and GhFD to form florigen activation complex that promoted flowering. The results provide a foundation for further studies on the regulatory mechanisms of flowering.

Download Full-text

Functional examination of lncRNAs in allotetraploid Gossypium hirsutum

BMC Genomics ◽

10.1186/s12864-021-07771-3 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Luyao Wang ◽

Jin Han ◽

Kening Lu ◽

Menglin Li ◽

Mengtao Gao ◽

...

Keyword(s):

Abiotic Stresses ◽

Expression Patterns ◽

Leaf Tissue ◽

Evolutionary Model ◽

Cotton Leaf ◽

Functional Screening ◽

Virus Induced Gene Silencing ◽

Individual Gene ◽

Cotton Species ◽

Allotetraploid Cotton

Abstract Background An evolutionary model using diploid and allotetraploid cotton species identified 80 % of non-coding transcripts in allotetraploid cotton as being uniquely activated in comparison with its diploid ancestors. The function of the lncRNAs activated in allotetraploid cotton remain largely unknown. Results We employed transcriptome analysis to examine the relationship between the lncRNAs and mRNAs of protein coding genes (PCGs) in cotton leaf tissue under abiotic stresses. LncRNA expression was preferentially associated with that of the flanking PCGs. Selected highly-expressed lncRNA candidates (n = 111) were subjected to a functional screening pilot test in which virus-induced gene silencing was integrated with abiotic stress treatment. From this low-throughput screen, we obtained candidate lncRNAs relating to plant height and tolerance to drought and other abiotic stresses. Conclusions Low-throughput screen is an effective method to find functional lncRNA for further study. LncRNAs were more active in abiotic stresses than PCG expression, especially temperature stress. LncRNA XLOC107738 may take a cis-regulatory role in response to environmental stimuli. The degree to which lncRNAs are constitutively expressed may impact expression patterns and functions on the individual gene level rather than in genome-wide aggregate.

Download Full-text