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

2021 ◽  
Vol 12 ◽  
Author(s):  
Tianyu Han ◽  
Wenlong Wu ◽  
Weilin Li

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.

2020 ◽  
Author(s):  
Tianyu Han ◽  
Zhixiang Yan ◽  
Wenlong Wu ◽  
Weilin Li

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.


2020 ◽  
Vol 145 (2) ◽  
pp. 131-140
Author(s):  
Xiaoying Dou ◽  
Jinrong Bai ◽  
Huan Wang ◽  
Ying Kong ◽  
Lixin Lang ◽  
...  

Anthocyanins are major pigments responsible for the color of lily (Lilium sp.) flowers. Anthocyanin synthesis is part of the flavonoid metabolic pathway. Numerous transcription factors, including R2R3-MYBs, basic helix-loop-helix (bHLH), and tryptophan–aspartic acid repeat (also known as WD40 or WD repeat) proteins, known to regulate flavonoid biosynthesis have been identified in various plant species. However, there is limited information available on WD repeat proteins in lilies. In this study, we identified a WD repeat gene in the Oriental hybrid lily ‘Sorbonne’ (Lilium hybrid WD repeat, LhWDR). LhWDR contains no introns, and has a 1100–base pair open reading frame, encoding a putative protein of 370 amino acids. LhWDR was found to be localized in the cytoplasm of transgenic Arabidopsis thaliana root cells. Expression patterns of LhWDR in different organs and at different periods of lily tepal growth revealed that the expression levels of this gene are closely associated with anthocyanin accumulation. A yeast two-hybrid assay demonstrated that full-length LhWDR interacts with the 420 N-terminal amino acids of Lilium hybrid bHLH2. Interestingly, overexpression of LhWDR in A. thaliana led to an upregulation of the dihydroflavonol 4-reductase gene, which is an important structural gene downstream of the anthocyanin pathway. These results indicate that the WD repeat protein LhWDR might interact with a bHLH transcription factor to regulate anthocyanin biosynthesis.


Horticulturae ◽  
2021 ◽  
Vol 7 (6) ◽  
pp. 123
Author(s):  
Haiying Yang ◽  
Changping Tian ◽  
Xiwen Li ◽  
Hansheng Gong ◽  
Aidi Zhang

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.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ming Li ◽  
Lei Sun ◽  
Hong Gu ◽  
Dawei Cheng ◽  
XiZhi Guo ◽  
...  

AbstractAs one of the largest transcription factor family, basic helix-loop-helix (bHLH) transcription factor family plays an important role in plant metabolism, physiology and growth. Berry color is one of the important factors that determine grape quality. However, the bHLH transcription factor family’s function in anthocyanin synthesis of grape berry has not been studied systematically. We identified 115 bHLH transcription factors in grape genome and phylogenetic analysis indicated that bHLH family could be classified into 25 subfamilies. First, we screened six candidate genes by bioinformatics analysis and expression analysis. We found one of the candidate genes VdbHLH037 belonged to III (f) subfamily and interacted with genes related to anthocyanin synthesis through phylogenetic analysis and interaction network prediction. Therefore, we speculated that VdbHLH037 participated in the anthocyanin synthesis process. To confirm this, we transiently expressed VdbHLH037 in grape and Arabidopsis transformation. Compared with the control, transgenic materials can accumulate more anthocyanins. These results provide a good base to study the function of the VdbHLH family in anthocyanin synthesis of grape berry.


2018 ◽  
Vol 44 (3) ◽  
pp. 289-298
Author(s):  
Bao-Jun Zhu ◽  
Qian Wang ◽  
Jing-Hui Wang ◽  
Lin-Lin Gao ◽  
Jing-Wen Zhang ◽  
...  

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.


2020 ◽  
Author(s):  
Nana Su ◽  
Ze Liu ◽  
Hui Chen ◽  
Mengyang Niu ◽  
Jin Cui

Abstract Background: The biosynthesis of anthocyanin in the hypocotyls of radish (Raphanus sativus L.) sprouts was enhanced by hemin in our preliminary experiments, but the underlying mechanism is unclear. Here, we found that NO (nitric oxide) exerted an essential role in Hemin-regulated anthocyanin biosynthesis, which was supported by the following results.Results: Hemin boosted anthocyanin as well as NO content. NO-scavenger cPTIO (carboxy-PTIO) significantly attenuated hemin-induced increase of anthocyanin content, transcripts of anthocyanin synthesis related genes and positive transcription factors, implying that NO played a prominent role during hemin-induced anthocyanin biosynthesis. Hemin specific inhibitor ZnPP (Zinc Protoporphyrin) strongly reduced anthocyanin content, while, NO donor SNP (Sodium Nitroprusside) addition considerably reversed this inhibition and by contrast, resulted in a significant increase in anthocyanin accumulation, closely paralleling the transcripts of structural genes and transcription factors. Moreover, NO content, NR (nitrate reductase) activity and expression level of NOA (nitric oxide associated factor) were up-regulated by Hemin. Conclusions:Those consequences indicated that NO might work downstream in Hemin-heightened anthocyanin accumulation in radish sprouts.


2019 ◽  
Vol 70 (12) ◽  
pp. 3111-3123 ◽  
Author(s):  
Lili Xiang ◽  
Xiaofen Liu ◽  
Heng Li ◽  
Xueren Yin ◽  
Donald Grierson ◽  
...  

Abstract ‘Jimba’, a well-known white flowered chrysanthemum cultivar, occasionally and spontaneously produces red colored petals under natural cultivation, but there is little information about the molecular regulatory mechanism underlying this process. We analysed the expression patterns of 91 MYB transcription factors in ‘Jimba’ and ‘Turning red Jimba’ and identified an R3 MYB, CmMYB#7, whose expression was significantly decreased in ‘Turning red Jimba’ compared with ‘Jimba’, and confirmed it is a passive repressor of anthocyanin biosynthesis. CmMYB#7 competed with CmMYB6, which together with CmbHLH2 is an essential component of the anthocyanin activation complex, for interaction with CmbHLH2 through the bHLH binding site in the R3 MYB domain. This reduced binding of the CmMYB6–CmbHLH2 complex and inhibited its ability to activate CmDFR and CmUFGT promoters. Moreover, using transient expression assays we demonstrated that changes in the expression of CmMYB#7 accounted for alterations in anthocyanin content. Taken together, our findings illustrate that CmMYB#7 is a negative regulator of anthocyanin biosynthesis in chrysanthemum.


Genome ◽  
2018 ◽  
Vol 61 (2) ◽  
pp. 111-120 ◽  
Author(s):  
Si-Won Jin ◽  
Md Abdur Rahim ◽  
Hoy-Taek Kim ◽  
Jong-In Park ◽  
Jong-Goo Kang ◽  
...  

Ornamental cabbage (Brassica oleracea var. acephala) is a winter-grown and important decorative plant of the family Brassicaceae, which displays an exceptional coloration in the central leaves of the rosette. Anthocyanins are the key determinant of the red, purple, and blue colors of vegetative and reproductive parts of many plant species including ornamental cabbage. Total anthocyanin content was measured spectrophotometrically, and the highest anthocyanin content was detected in the red followed by light-red and white ornamental cabbage lines. Anthocyanin biosynthesis is controlled by members of three different transcription factor (TF) families, such as MYB, basic helix-loop-helix (bHLH), and WD40 repeats (WDR), which function as a MBW complex. We identified three MYB, six bHLH, and one WDR TFs that regulate anthocyanin biosynthesis in ornamental cabbage. The expression of the regulatory and biosynthetic genes for anthocyanin synthesis was determined by qPCR. The tested structural genes of the anthocyanin pathway were shown to be up-regulated in the red followed by light-red ornamental cabbage lines; however, the expression levels of the late biosynthetic genes were barely detected in the white ornamental cabbage lines. Among the regulatory genes, BoPAP2 (MYB), BoTT8, BoEGL3.1, and BoMYC1.2 (bHLH), and BoTTG1 (WDR) were identified as candidates for the regulation of anthocyanin biosynthesis. This work could be useful for the breeding of novel colorful ornamental cabbage cultivars.


Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2721
Author(s):  
Chao Tan ◽  
Huilei Qiao ◽  
Ming Ma ◽  
Xue Wang ◽  
Yunyun Tian ◽  
...  

The basic helix-loop-helix (bHLH) transcription factor family is one of the largest transcription factor families in plants and plays crucial roles in plant development. Melon is an important horticultural plant as well as an attractive model plant for studying fruit ripening. However, the bHLH gene family of melon has not yet been identified, and its functions in fruit growth and ripening are seldom researched. In this study, 118 bHLH genes were identified in the melon genome. These CmbHLH genes were unevenly distributed on chromosomes 1 to 12, and five CmbHLHs were tandem repeat on chromosomes 4 and 8. There were 13 intron distribution patterns among the CmbHLH genes. Phylogenetic analysis illustrated that these CmbHLHs could be classified into 16 subfamilies. Expression patterns of the CmbHLH genes were studied using transcriptome data. Tissue specific expression of the CmbHLH32 gene was analysed by quantitative RT-PCR. The results showed that the CmbHLH32 gene was highly expressed in female flower and early developmental stage fruit. Transgenic melon lines overexpressing CmbHLH32 were generated, and overexpression of CmbHLH32 resulted in early fruit ripening compared to wild type. The CmbHLH transcription factor family was identified and analysed for the first time in melon, and overexpression of CmbHLH32 affected the ripening time of melon fruit. These findings laid a foundation for further study on the role of bHLH family members in the growth and development of melon.


2018 ◽  
Vol 45 (No. 1) ◽  
pp. 1-10 ◽  
Author(s):  
Gijing Zhang ◽  
Tong Li ◽  
Lijie zhang ◽  
Wenxuan dong ◽  
Aide Wang

Plant-specific NAC transcription factors (TFs) play crucial roles in various pathways related to the stress response. However, to date, little information regarding NAC gene regulation during fruit ripening is available for the apple (Malus domestica). Here, we report that 13 out of 182 MdNAC genes were differentially expressed during the stages of fruit growth and ripening. Sequence analysis indicates that these 13 MdNAC genes harbour distinct structures and potentially diverse functions. The expression of both MdNAC1a and MdNAC78 was repressed by ethylene and induced by 1-MCP during storage. MdNAC2, MdNAC26, MdNAC41, MdNAC57, MdNAC80, MdNAC91, MdNAC119 and MdNAC141 were up-regulated by ethylene and their transcription mirrored ethylene production rates during storage. MdNAC1, MdNAC16 and MdNAC32 did not respond to 1-MCP exposure. Additionally, the 13 MdNAC genes identified displayed differential tissue-specific expression patterns. These results suggest that NAC TFs play an important role in the regulation of apple development via both ethylene-dependent and -independent mechanisms.


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