scholarly journals Functional Characterization and in Silico Analysis of Phytoene Synthase Family Genes Responsible for Carotenoid Biosynthesis in Watermelon (Citrullus lanatus L.)

Agronomy ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 1077 ◽  
Author(s):  
Chuan Wu ◽  
Lei Sun ◽  
Yuanzuo Lv ◽  
Haonan Cui ◽  
Xuezheng Wang ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Fruit Ripening ◽  
Developmental Stages ◽  
Citrullus Lanatus ◽  
Dominant Role ◽  
Functional Characterization ◽  
In Silico Analysis ◽  
Carotenoid Biosynthesis ◽  
Promoter Sequence ◽  
Phytoene Synthase

Carotenoids are the main pigments in watermelon (Citrullus lanatus L.) fruit and contribute to its aesthetic and nutritional value. Phytoene synthase (PSY) is reported to be the first rate-limiting enzyme in carotenogenesis and controls the carotenoid flux. This study aimed to identify PSY genes responsible for carotenoid biosynthesis in the red-fleshed watermelon cultivar LSW-177. The PSY gene members ClPSY1, ClPSY2 and ClPSY3 were characterized and their catalytic activities were displayed in the heterologous complementation assay. The transcript levels of ClPSY genes at the different developmental stages of LSW-177 fruit and the promoter sequence of ClPSY1 were also analyzed. Transcription factors involved in regulating the ClPSY1 expression were scanned with previous RNA-seq data of the different stages during fruit ripening. Results showed that the PSY proteins from watermelon LSW-177 contained the conserved PSY domains and exhibited the ability to condense GGPP into phytoene in E. coli. ClPSY1 is the dominant carotenogenic gene during fruit ripening; and can be induced by light and hormones. Furthermore, Cla013914 and Cla007950 that, respectively encode the transcription factors WD40-like protein and bZIP, likely upregulate ClPSY1 during fruit ripening. In conclusion, ClPSY1 play a dominant role in carotenoid biosynthesis during watermelon fruit ripening and is regulated by complex light and hormone-responsive networks.

scholarly journals Transcriptome Regulation of Carotenoids in Five Flesh-Colored Watermelon (Citrullus lanatus)

2020 ◽  
Author(s):  
Pingli Yuan ◽  
Muhammad Jawad Umer ◽  
Nan He ◽  
Shengjie Zhao ◽  
Xuqiang Lu ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Molecular Mechanisms ◽  
Citrullus Lanatus ◽  
Carotenoid Biosynthesis ◽  
Flesh Color ◽  
Hub Genes ◽  
Color Formation ◽  
Transcriptome Regulation ◽  
Yellow Flesh ◽  
Carotenoid Biosynthesis Pathway

Abstract Background: Fruit flesh color in watermelon (Citrullus lanatus) is a great index for evaluation of the appearance quality and a key contributor influencing consumers preferences, but the molecular mechanisms of this intricate trait remain largely unknown. Here, the carotenoids and transcriptome dynamics during fruit development in watermelon cultivars with 5 different flesh colors were analyzed.Results: A total of 13 carotenoids and 16,781 differentially expressed genes (DEGs) including 1,295 transcription factors (TFs) were detected during the development of five watermelon genotypes. A number of structural genes and transcription factors were found to be involved in the carotenoid biosynthesis pathway using comparative transcriptome analysis. Furthermore, we performed weighted gene co-expression network analysis and predicted hub genes in 6 main modules determining carotenoids contents. Cla018406 (a Chaperone protein dnaJ-like protein) maybe a candidate gene for β-carotene and highly expressed in orange flesh colored fruit. Cla007686 (a zinc finger CCCH domain-containing protein) was highly expressed in the red color watermelon, maybe a key regulator for lycopene accumulation. Cla003760 (merbrane protein) and Cla007686 (photosystenI reaction center subunit II) are predicted to be hub genes and play an important role in yellow flesh color formation.Conclusions: These results provide an important resource for dissecting the molecular basis and candidate genes governing flesh color formation in watermelon fruit.

scholarly journals Screening and Identification of Transcription Factors Potentially Regulating Foxl2 Expression in Chlamys farreri Ovary

Biology ◽  
2022 ◽  
Vol 11 (1) ◽  
pp. 113
Author(s):  
Shutong Fan ◽  
Xixi Li ◽  
Siyu Lin ◽  
Yunpeng Li ◽  
Huixin Ma ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Ovarian Function ◽  
Developmental Stages ◽  
Promoter Sequence ◽  
Luciferase Reporter ◽  
Ovary Development ◽  
Regulation Mechanism ◽  
Specific Expression ◽  
Screening And Identification ◽  
Specific Regulation

Foxl2 is an evolutionarily conserved female sex gene, which is specifically expressed in the ovary and mainly involved in oogenesis and ovarian function maintenance. However, little is known about the mechanism that regulates Foxl2 specific expression during the ovary development. In the present study, we constructed the gonadal yeast one-hybrid (Y1H) library of Chlamysfarreri with ovaries and testes at different developmental stages using the Gateway technology. The library capacity was more than 1.36 × 107 CFU, and the length of the inserted fragment was 0.75 Kb~2 Kb, which fully met the demand of yeast library screening. The highly transcriptional activity promoter sequence of C. farreri Foxl2 (Cf-Foxl2) was determined at −1000~−616 bp by dual-luciferase reporter (DLR) assay and was used as bait to screen possible transcription factors from the Y1H library. Eleven candidate factors, including five unannotated factors, were selected based on Y1H as well as their expressional differences between ovaries and testes and were verified for the first time to be involved in the transcriptional regulation of Cf-Foxl2 by RT-qPCR and DLR. Our findings provided valuable data for further studying the specific regulation mechanism of Foxl2 in the ovary.

scholarly journals Transcriptome regulation of carotenoids in five flesh-colored watermelon (Citrullus lanatus)

2021 ◽  
Author(s):  
Pingli Yuan ◽  
Muhammad Jawad Umer ◽  
Nan He ◽  
Shengjie Zhao ◽  
Xuqiang Lu ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Candidate Genes ◽  
Differentially Expressed Genes ◽  
Fruit Development ◽  
Citrullus Lanatus ◽  
Carotenoid Biosynthesis ◽  
Differentially Expressed ◽  
Flesh Color ◽  
Hub Genes ◽  
Data Resource

Abstract Background: Fruit flesh color in watermelon (Citrullus lanatus) is a great index for evaluation of the appearance quality and a key contributor influencing consumers' preferences, but the molecular mechanism of this intricate trait remain largely unknown. Here, the carotenoids and transcriptome dynamics during the fruit development of cultivated watermelon with five different flesh colors were analyzed.Results: A total of 13 carotenoids and 16781 differentially expressed genes (DEGs) including 1295 transcription factors (TFs) were detected in five watermelon genotypes during the fruit development. The comprehensive accumulation patterns of carotenoids were closely related to flesh color. A number of potential structural genes and transcription factors were found to be associated with the carotenoid biosynthesis pathway using comparative transcriptome analysis. The differentially expressed genes were divided into six subclusters and distributed in different GO terms and metabolic pathways. Furthermore, we performed weighted gene co-expression network analysis and predicted hub genes in six main modules determining carotenoid contents. Cla018406 (a chaperone protein dnaJ-like protein) may be a candidate gene for β-carotene accumulation and highly expressed in orange flesh-colored fruit. Cla007686 (a zinc finger CCCH domain-containing protein) was highly expressed in the red flesh-colored watermelon, maybe a key regulator of lycopene accumulation. Cla003760 (membrane protein) and Cla021635 (photosystem I reaction center subunit II) were predicted to be hub genes and may play an essential role in yellow flesh formation.Conclusions: The composition and contents of carotenoid in five watermelon genotypes vary greatly. A series of candidate genes were revealed through combined analysis of metabolites and transcriptome. These results provide an important data resource for dissecting the candidate genes and molecular basis governing flesh color formation in watermelon fruit.

scholarly journals Characterization of BoaCRTISO Reveals Its Role in Carotenoid Biosynthesis in Chinese Kale

2021 ◽  
Vol 12 ◽  
Author(s):  
Min Jiang ◽  
Fen Zhang ◽  
Qiao Yuan ◽  
Peixing Lin ◽  
Hao Zheng ◽  
...  
Keyword(s):  
Acid Treatment ◽  
Biosynthetic Pathway ◽  
Developmental Stages ◽  
Functional Characterization ◽  
Carotenoid Biosynthesis ◽  
Virus Induced Gene Silencing ◽  
Strong Light ◽  
Organic Pigments ◽  
Chinese Kale

Carotenoids are organic pigments that play an important role in both plant coloration and human health; they are a critical subject in molecular breeding due to growing demand for natural molecules in both food and medicine. In this study, we focus upon characterizing BoaCRTISO, the carotenoid isomerase gene before the branch of the carotenoid biosynthetic pathway, which is expressed in all organs and developmental stages of Chinese kale, and BoaCRTISO, which is located in the chloroplast. The expression of BoaCRTISO is induced by strong light, red and blue combined light, and gibberellic acid treatment, but it is suppressed by darkness and abscisic acid treatment. We obtained BoaCRTISO-silenced plants via virus-induced gene silencing technology, and the silence efficiencies ranged from 52 to 77%. The expressions of most carotenoid and chlorophyll biosynthetic genes in BoaCRTISO-silenced plants were downregulated, and the contents of carotenoids and chlorophyll were reduced. Meanwhile, BoaCRTISO-silenced plants exhibited phenotypes of yellowing leaves and inhibited growth. This functional characterization of BoaCRTISO provides insight for the biosynthesis and regulation of carotenoid in Chinese kale.

scholarly journals Transcriptome regulation of carotenoids in five flesh-colored watermelons (Citrullus lanatus)

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Pingli Yuan ◽  
Muhammad Jawad Umer ◽  
Nan He ◽  
Shengjie Zhao ◽  
Xuqiang Lu ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Candidate Genes ◽  
Differentially Expressed Genes ◽  
Fruit Development ◽  
Citrullus Lanatus ◽  
Carotenoid Biosynthesis ◽  
Differentially Expressed ◽  
Flesh Color ◽  
Hub Genes ◽  
Data Resource

Abstract Background Fruit flesh color in watermelon (Citrullus lanatus) is a great index for evaluating the appearance quality and a key contributor influencing consumers’ preferences. But the molecular mechanism of this intricate trait remains largely unknown. Here, the carotenoids and transcriptome dynamics during the fruit development of cultivated watermelon with five different flesh colors were analyzed. Results A total of 13 carotenoids and 16,781 differentially expressed genes (DEGs), including 1295 transcription factors (TFs), were detected in five watermelon genotypes during the fruit development. The comprehensive accumulation patterns of carotenoids were closely related to flesh color. A number of potential structural genes and transcription factors were found to be associated with the carotenoid biosynthesis pathway using comparative transcriptome analysis. The differentially expressed genes were divided into six subclusters and distributed in different GO terms and metabolic pathways. Furthermore, we performed weighted gene co-expression network analysis and predicted the hub genes in six main modules determining carotenoid contents. Cla018406 (a chaperone protein dnaJ-like protein) may be a candidate gene for β-carotene accumulation and highly expressed in orange flesh-colored fruit. Cla007686 (a zinc finger CCCH domain-containing protein) was highly expressed in the red flesh-colored watermelon, maybe a key regulator of lycopene accumulation. Cla003760 (membrane protein) and Cla021635 (photosystem I reaction center subunit II) were predicted to be the hub genes and may play an essential role in yellow flesh formation. Conclusions The composition and contents of carotenoids in five watermelon genotypes vary greatly. A series of candidate genes were revealed through combined analysis of metabolites and transcriptome. These results provide an important data resource for dissecting candidate genes and molecular basis governing flesh color formation in watermelon fruit.

scholarly journals Genome-wide identification, classification, and expression analysis of the HSF gene family in pineapple (Ananas comosus)

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11329
Author(s):  
Lulu Wang ◽  
Yanhui Liu ◽  
Mengnan Chai ◽  
Huihuang Chen ◽  
Mohammad Aslam ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Gene Family ◽  
Plant Development ◽  
Developmental Stages ◽  
Stress Hormones ◽  
Functional Characterization ◽  
Ananas Comosus ◽  
Group A ◽  
Group B ◽  
Regulatory Functions

Transcription factors (TFs), such as heat shock transcription factors (HSFs), usually play critical regulatory functions in plant development, growth, and response to environmental cues. However, no HSFs have been characterized in pineapple thus far. Here, we identified 22 AcHSF genes from the pineapple genome. Gene structure, motifs, and phylogenetic analysis showed that AcHSF families were distinctly grouped into three subfamilies (12 in Group A, seven in Group B, and four in Group C). The AcHSF promoters contained various cis-elements associated with stress, hormones, and plant development processes, for instance, STRE, WRKY, and ABRE binding sites. The majority of HSFs were expressed in diverse pineapple tissues and developmental stages. The expression of AcHSF-B4b/AcHSF-B4c and AcHSF-A7b/AcHSF-A1c were enriched in the ovules and fruits, respectively. Six genes (AcHSF-A1a , AcHSF-A2, AcHSF-A9a, AcHSF-B1a, AcHSF-B2a, and AcHSF-C1a) were transcriptionally modified by cold, heat, and ABA. Our results provide an overview and lay the foundation for future functional characterization of the pineapple HSF gene family.

scholarly journals Genome wide analysis of kinesin gene family in Citrullus lanatus reveals an essential role in early fruit development

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Shujuan Tian ◽  
Jiao Jiang ◽  
Guo-qi Xu ◽  
Tan Wang ◽  
Qiyan Liu ◽  
...  
Keyword(s):  
Gene Family ◽  
Plant Hormones ◽  
Fruit Development ◽  
Developmental Stages ◽  
Profile Analysis ◽  
Citrullus Lanatus ◽  
Binding Motif ◽  
Distribution Analysis ◽  
Motif Analysis ◽  
Expression Profile Analysis

Abstract Background Kinesin (KIN) as a motor protein is a versatile nano-machine and involved in diverse essential processes in plant growth and development. However, the kinesin gene family has not been identified in watermelon, a valued and nutritious fruit, and yet their functions have not been characterized. Especially, their involvement in early fruit development, which directly determines the size, shape, yield and quality of the watermelon fruit, remains unclear. Results In this study, we performed a whole-genome investigation and comprehensive analysis of kinesin genes in C. lanatus. In total, 48 kinesins were identified and categorized into 10 kinesin subfamilies groups based on phylogenetic analysis. Their uneven distribution on 11 chromosomes was revealed by distribution analysis. Conserved motif analysis showed that the ATP-binding motif of kinesins was conserved within all subfamilies, but not the microtubule-binding motif. 10 segmental duplication pairs genes were detected by the syntenic and phylogenetic approaches, which showed the expansion of the kinesin gene family in C. lanatus genome during evolution. Moreover, 5 ClKINs genes are specifically and abundantly expressed in early fruit developmental stages according to comprehensive expression profile analysis, implying their critical regulatory roles during early fruit development. Our data also demonstrated that the majority of kinesin genes were responsive to plant hormones, revealing their potential involvement in the signaling pathways of plant hormones. Conclusions Kinesin gene family in watermelon was comprehensively analyzed in this study, which establishes a foundation for further functional investigation of C. lanatus kinesin genes and provides novel insights into their biological functions. In addition, these results also provide useful information for understanding the relationship between plant hormone and kinesin genes in C. lanatus.

scholarly journals Regulatory Network Analysis in Estradiol-Treated Human Endothelial Cells

2021 ◽  
Vol 22 (15) ◽  
pp. 8193
Author(s):  
Daniel Pérez-Cremades ◽  
Ana B. Paes ◽  
Xavier Vidal-Gómez ◽  
Ana Mompeón ◽  
Carlos Hermenegildo ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Transcription Factor ◽  
Transcription Factors ◽  
Regulatory Network ◽  
Mirna Target ◽  
Target Genes ◽  
Functional Characterization ◽  
Human Endothelial Cells ◽  
Mrna Targets ◽  
Canonical Pathways

Background/Aims: Estrogen has been reported to have beneficial effects on vascular biology through direct actions on endothelium. Together with transcription factors, miRNAs are the major drivers of gene expression and signaling networks. The objective of this study was to identify a comprehensive regulatory network (miRNA-transcription factor-downstream genes) that controls the transcriptomic changes observed in endothelial cells exposed to estradiol. Methods: miRNA/mRNA interactions were assembled using our previous microarray data of human umbilical vein endothelial cells (HUVEC) treated with 17β-estradiol (E2) (1 nmol/L, 24 h). miRNA–mRNA pairings and their associated canonical pathways were determined using Ingenuity Pathway Analysis software. Transcription factors were identified among the miRNA-regulated genes. Transcription factor downstream target genes were predicted by consensus transcription factor binding sites in the promoter region of E2-regulated genes by using JASPAR and TRANSFAC tools in Enrichr software. Results: miRNA–target pairings were filtered by using differentially expressed miRNAs and mRNAs characterized by a regulatory relationship according to miRNA target prediction databases. The analysis identified 588 miRNA–target interactions between 102 miRNAs and 588 targets. Specifically, 63 upregulated miRNAs interacted with 295 downregulated targets, while 39 downregulated miRNAs were paired with 293 upregulated mRNA targets. Functional characterization of miRNA/mRNA association analysis highlighted hypoxia signaling, integrin, ephrin receptor signaling and regulation of actin-based motility by Rho among the canonical pathways regulated by E2 in HUVEC. Transcription factors and downstream genes analysis revealed eight networks, including those mediated by JUN and REPIN1, which are associated with cadherin binding and cell adhesion molecule binding pathways. Conclusion: This study identifies regulatory networks obtained by integrative microarray analysis and provides additional insights into the way estradiol could regulate endothelial function in human endothelial cells.

scholarly journals TBP and SNAP50 transcription factors bind specifically to the Pr77 promoter sequence from trypanosomatid non-LTR retrotransposons

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Francisco Macías ◽  
Raquel Afonso-Lehmann ◽  
Patricia E. Carreira ◽  
M. Carmen Thomas
Keyword(s):  
Transcription Factors ◽  
Protein Interactions ◽  
Dna Sequences ◽  
Direct Interaction ◽  
Promoter Sequence ◽  
Nuclear Proteins ◽  
Hill Coefficient ◽  
Small Nuclear Rna ◽  
Mobile Dna ◽  
Mobility Shift

Abstract Background Trypanosomatid genomes are colonized by active and inactive mobile DNA elements, such as LINE, SINE-like, SIDER and DIRE retrotransposons. These elements all share a 77-nucleotide-long sequence at their 5′ ends, known as Pr77, which activates transcription, thereby generating abundant unspliced and translatable transcripts. However, transcription factors that mediates this process have still not been reported. Methods TATA-binding protein (TBP) and small nuclear RNA-activating protein 50 kDa (SNAP50) recombinant proteins and specific antibodies raised against them were generated. Protein capture assay, electrophoretic mobility-shift assays (EMSA) and EMSA competition assays carried out using these proteins and nuclear proteins of the parasite together to specific DNA sequences used as probes allowed detecting direct interaction of these transcription factors to Pr77 sequence. Results This study identified TBP and SNAP50 as part of the DNA-protein complex formed by the Pr77 promoter sequence and nuclear proteins of Trypanosoma cruzi. TBP establishes direct and specific contact with the Pr77 sequence, where the DPE and DPE downstream regions are docking sites with preferential binding. TBP binds cooperatively (Hill coefficient = 1.67) to Pr77 and to both strands of the Pr77 sequence, while the conformation of this highly structured sequence is not involved in TBP binding. Direct binding of SNAP50 to the Pr77 sequence is weak and may be mediated by protein–protein interactions through other trypanosomatid nuclear proteins. Conclusions Identification of the transcription factors that mediate Pr77 transcription may help to elucidate how these retrotransposons are mobilized within the trypanosomatid genomes and their roles in gene regulation processes in this human parasite. Graphic abstract

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