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

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.

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Transcriptional Profiling of Methyltransferase Genes during Growth of Methanosarcina mazei on Trimethylamine

Journal of Bacteriology ◽

10.1128/jb.00420-09 ◽

2009 ◽

Vol 191 (16) ◽

pp. 5108-5115 ◽

Cited By ~ 17

Author(s):

Christian Krätzer ◽

Paul Carini ◽

Raymond Hovey ◽

Uwe Deppenmeier

Keyword(s):

Transcriptional Profiling ◽

Expression Patterns ◽

Pcr Analysis ◽

Exponential Growth Phase ◽

Methane Formation ◽

Rt Pcr ◽

Transcript Levels ◽

Methanosarcina Mazei ◽

Depth Analysis ◽

Genes Encoding

ABSTRACT The genomic expression patterns of Methanosarcina mazei growing with trimethylamine were measured in comparison to those of cells grown with methanol. We identified a total of 72 genes with either an increased level (49 genes) or a decreased level (23 genes) of mRNA during growth on trimethylamine with methanol-grown cells as the control. Major differences in transcript levels were observed for the mta, mtb, mtt, and mtm genes, which encode enzymes involved in methane formation from methanol and trimethylamine, respectively. Other differences in mRNA abundance were found for genes encoding enzymes involved in isopentenyl pyrophosphate synthesis and in the formation of aromatic amino acids, as well as a number of proteins with unknown functions. The results were verified by in-depth analysis of methyltransferase genes using specific primers for real-time quantitative reverse transcription-PCR (RT-PCR). The monitored transcript levels of genes encoding corrinoid proteins involved in methyl group transfer from methylated C1 compounds (mtaC, mtbC, mttC, and mtmC) indicated increased amounts of mRNA from the mtaBC1, mtaBC2, and mtaBC3 operons in methanol-grown cells, whereas mRNA of the mtb1-mtt1 operon was found in high concentrations during trimethylamine consumption. The genes of the mtb1-mtt1 operon encode methyltransferases that are responsible for sequential demethylation of trimethylamine. The analysis of product formation of trimethylamine-grown cells at different optical densities revealed that large amounts of dimethylamine and monomethylamine were excreted into the medium. The intermediate compounds were consumed only in the very late exponential growth phase. RT-PCR analysis of key genes involved in methanogenesis led to the conclusion that M. mazei is able to adapt to changing trimethylamine concentrations and the consumption of intermediate compounds. Hence, we assume that the organism possesses a regulatory network for optimal substrate utilization.

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Time-Series Transcriptome Sheds Light on Anthocyanin Metabolism and Species Diversification Between two Lonicera Japonica Thunb Cultivars

10.21203/rs.3.rs-104094/v1 ◽

2020 ◽

Author(s):

Jianjun Li ◽

Chenglin Ye ◽

Jingxiao Ma ◽

Ting Cheng ◽

Yan Lv ◽

...

Keyword(s):

Flower Color ◽

Carotenoid Biosynthesis ◽

Lonicera Japonica ◽

Pcr Analysis ◽

Anthocyanin Synthesis ◽

New Variety ◽

Color Development ◽

Phenylpropanoid Biosynthesis ◽

New Varieties ◽

Three Stages

Abstract Background：‘Yujin 2’ is new variety of Lonicera japonica Thunb and its flower color can change from red to yellow; hence, it is a good model for investigating flower color development mechanisms. Results：High throughput transcriptome sequencing of seven flower development stages of Yujin No.2 was carried out, and 133,487 unigenes were annotated, among which 73,088 were differentially expressed. Then the real-time PCR analysis was carried out. Further, the number of up-regulated DEGs was higher than those that were down-regulated. Of these annotated DEGs, plant hormone signal transduction, phenylpropanoid biosynthesis, and flavonoid biosynthesis were active throughout the flowering process during each stage, whereas carotenoid biosynthesis was inactive in the S1-6 stages. Furthermore, phenylalanine synthesis was enhanced in the S1 phase; however, anthocyanin synthesis was weakened in the S5 and S6 phases, which may be consistent with the changes in petal color of ‘Yujin 2’ from red (S1) to white (S5) and gold (S6). The results showed that 114 unigenes were associated with anthocyanin metabolism, and 72 were significantly upregulated or downregulated. According to the analysis of TFs in anthocyanin metabolism, we obtained 47 transcription factors, which belonged to 18 families. The LjDFR, LjABCB1, LjMYC6, LjDDB2, and LjANS genes rapidly increased during the first three stages. However, only LjF3'5'H expression was significantly down-regulated at S5, which was consistent with anthocyanin accumulation. Conclusions：This study developed a transcriptome profile of flower color generation for L. japonica as well as annotated unigene sets of seven anthesis phases, thereby providing possibilities for improving the germplasm of L. japonica with genetic engineering technologies and cultivating new varieties of different colors.

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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.

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Comparative Transcriptome Analysis Reveals Effects of Exogenous Hematin on Anthocyanin Biosynthesis during Strawberry Fruit Ripening

International Journal of Genomics ◽

10.1155/2016/6762731 ◽

2016 ◽

Vol 2016 ◽

pp. 1-14 ◽

Cited By ~ 5

Author(s):

Yi Li ◽

Huayin Li ◽

Fengde Wang ◽

Jingjuan Li ◽

Yihui Zhang ◽

...

Keyword(s):

Transcriptome Analysis ◽

Regulatory Networks ◽

Anthocyanin Biosynthesis ◽

Anthocyanin Synthesis ◽

Control Group ◽

Anthocyanin Content ◽

Color Development ◽

Strawberry Fruit Ripening ◽

Positive Effect

Anthocyanin in strawberries has a positive effect on fruit coloration. In this study, the role of exogenous hematin on anthocyanin biosynthesis was investigated. Our result showed that the white stage of strawberries treated with exogenous hematin had higher anthocyanin content, compared to the control group. Among all treatments, 5 μM of hematin was the optimal condition to promote color development. In order to explore the molecular mechanism of fruit coloring regulated by hematin, transcriptomes in the hematin- and non-hematin-treated fruit were analyzed. A large number of differentially expressed genes (DEGs) were identified in regulating anthocyanin synthesis, including the DEGs involved in anthocyanin biosynthesis, hormone signaling transduction, phytochrome signaling, starch and sucrose degradation, and transcriptional pathways. These regulatory networks may play an important role in regulating the color process of strawberries treated with hematin. In summary, exogenous hematin could promote fruit coloring by increasing anthocyanin content in the white stage of strawberries. Furthermore, transcriptome analysis suggests that hematin-promoted fruit coloring occurs through multiple related metabolic pathways, which provides valuable information for regulating fruit color via anthocyanin biosynthesis in strawberries.

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Transcriptional Inactivation of p53, Bax, Bcl-2 and Mdm2 Correlates with Malignant Transformation of the Uterine Cervix

The International Journal of Biological Markers ◽

10.1177/172460080502000104 ◽

2005 ◽

Vol 20 (1) ◽

pp. 18-27 ◽

Cited By ~ 5

Author(s):

G. Soufla ◽

S. Baritaki ◽

S. Sifakis ◽

A. Zafiropoulos ◽

D.A. Spandidos

Keyword(s):

Mrna Expression ◽

Malignant Transformation ◽

Uterine Cervix ◽

Expression Patterns ◽

Cellular Transformation ◽

Pcr Analysis ◽

Mrna Levels ◽

Cervical Lesions ◽

Transcript Levels ◽

Mdm2 Mrna

Deregulation of the apoptotic machinery plays a major role in cell death, cellular transformation and cancer. p53, Bcl-2, Bcl-XL, Bax and Mdm2 mRNA expression patterns were evaluated in tissue samples with cervical intraepithelial neoplasia (CIN) and cervical cancer compared to those of normal cervical tissues, and correlated with the underlying cervical lesions. Transcript levels of the above genes were assessed by RT-PCR analysis in a total of 44 cervical specimens. p53, Bcl-2, Bax and Mdm2 transcript levels were significantly different in the normal, CIN and cancer specimen groups (p=0.003, p=0.009, p=0.040 and p=0.001, respectively). Specifically, p53, Bax and Bcl-2 exhibited substantially lower transcript levels in CIN lesions compared to controls, whereas Bax mRNA levels showed a significant decrease in cancer compared to normal specimens. Mdm2 mRNA expression was considerably lower in cancer than in CIN lesions or normal cervix. High-grade squamous intraepithelial lesions exhibited lower p53 and Bcl-2 mRNA levels than controls (p=0.002, p=0.016). Coexpression analysis revealed more correlations between the above apoptosis-related molecules in normal tissues compared to CIN or cancer specimens. p53 showed significant coexpression with Bax, Bcl-2 and Mdm2 (p=0.040, p=0.013 and p=0.015, respectively) in normal cervical specimens. Bax and Bcl-XL mRNA expression was negatively correlated. Mdm2 transcriptional levels correlated significantly with those of Bax, Bcl-XL and Bcl-2. Our findings show that p53, Bax, Bcl-2 and Mdm2 mRNA expression levels correlate with the malignant transformation of the uterine cervix. mRNA coexpression patterns of the members of the pro- and anti-apoptotic family examined in cervical carcinogenesis were found to be disrupted in CIN and cancer, as already demonstrated at the protein level.

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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.

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197 Effect of STC-4771, An Intermediate of ABA Synthesis, on the Anthocyanin Accumulation in Grapes

HortScience ◽

10.21273/hortsci.35.3.425a ◽

2000 ◽

Vol 35 (3) ◽

pp. 425A-425

Author(s):

W.S. Lee ◽

J.C. Lee ◽

Y.S. Hwang

Keyword(s):

Anthocyanin Synthesis ◽

Anthocyanin Content ◽

Internal Factors ◽

External Application ◽

Grape Berries ◽

Color Development ◽

Agricultural Use ◽

Favorable Conditions

The coloration of grape berries depends on the anthocyanin synthesis during maturation. The quality of berries is often decreased due to the poor color development when berries are grown under unfavorable environments and/or inadequate internal factors are involved. It has been well-known that the level of ABA at ripening is closely associated with anthocyanin synthesis; thus, the external application of ABA results in the increase of anthocyanin content even in berries grown under favorable conditions. However, the agricultural use of natural ABA is not possible because of high prices. This experiment was conducted to study the potential of STC-4771 as a substitute for ABA. The effect of STC-4771 was studied in `Kyoho', `Pione', and `Delaware' grapes. Chemicals were applied when ≈10% of berries in a cluster were colored. In `Kyoho', anthocyanin synthesis was enhanced at a concentration of 100 mg/L and there was a trend in color enhancement in `Pione', regardless of treatment concentration, between 10 to 40 mg/L. However, no clear effect was found in `Delaware' at 50 to 100 mg/L. In an in vitro experiment, anthocyanin was only increased when an adequate amount of sucrose (0.6 m) was added in the incubation medium under light. Natural ABA effectively increased the anthocyanin content of berry segments even under shading condition through four bagging materials, but no effect was confirmed in STC treatment.

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Influence of Phostrade Ca on Color Development and Anthocyanin Content of ‘Braeburn’ Apple (Malus domestica Borkh.)

HortScience ◽

10.21273/hortsci.48.2.193 ◽

2013 ◽

Vol 48 (2) ◽

pp. 193-199 ◽

Cited By ~ 10

Author(s):

Jan Bizjak ◽

Nika Weber ◽

Maja Mikulic-Petkovsek ◽

Ana Slatnar ◽

Franci Stampar ◽

...

Keyword(s):

Hydroxycinnamic Acids ◽

Anthocyanin Synthesis ◽

Total Phenolic ◽

Anthocyanin Content ◽

Total Sugars ◽

Primary And Secondary Metabolites ◽

Color Development ◽

Commercial Harvest ◽

Significant Difference ◽

High Concentrations

The influence of two foliar applications of Phostrade Ca, which contains high concentrations of phosphorus and minor amounts of calcium and nitrogen, on color development and selected primary and secondary metabolites was investigated during advanced maturation of ‘Braeburn’ apple. Changes of hydroxycinnamic acids, flavanols, dihydrochalcones, flavonols, and anthocyanins were monitored six times during the advanced ripening until technological maturity of the fruits. Additionally, the changes in the chromatic values a*, h°, and the lightness coefficient L* were recorded weekly. The colorimetric parameters showed a significant difference in the intensity of red coloration between the treated and untreated apples. Spraying with Phostrade Ca also resulted in a significant increase in most individual sugars, total sugars, and concentration of anthocyanins and flavonols. Moreover, the amount of phosphorus (P) in the treated leaves was increased. However, the total phenolic content and accumulation of other classes of flavonoids such as hydroxycinnamic acids, flavonols, and dihydrochalcones were not influenced. Phostrade Ca treatment significantly increased dihydroflavonol 4-reductase (DFR) and slightly flavanone-3-hydroxylase (FHT) activity, which were correlated with anthocyanin synthesis but had no effect on phenylalanine ammonia lyase (PAL) and chalcone synthase/chalcone isomerase (CHS/CHI) activity. The results indicate that two foliar applications of Phostrade Ca late in the growing season represent an effective way to improve the color of ‘Braeburn’ apples at commercial harvest.

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Characterization of Three Chalcone Synthase-like Genes in Dianthus Chinensis

10.21203/rs.3.rs-159109/v1 ◽

2021 ◽

Author(s):

Jia Liu ◽

Xi-Long Hao ◽

Xue-Qin He

Keyword(s):

Chalcone Synthase ◽

Expression Patterns ◽

Tobacco Rattle Virus ◽

Transcript Level ◽

Close Relation ◽

Amino Acid Sequences ◽

Anthocyanin Synthesis ◽

Anthocyanin Content ◽

Virus Induced Gene Silencing ◽

Floral Bud

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.

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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.

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