scholarly journals Isolation and Functional Analysis of Genes Involved in Polyacylated Anthocyanin Biosynthesis in Blue Senecio cruentus

2021 ◽  
Vol 12 ◽  
Author(s):  
Chenfei Lu ◽  
Yajun Li ◽  
Yumeng Cui ◽  
Jiangshan Ren ◽  
Fangting Qi ◽  
...  
Keyword(s):  
Molecular Breeding ◽  
Higher Plants ◽  
Anthocyanin Biosynthesis ◽  
Functional Characterization ◽  
Glycoside Hydrolase Family ◽  
Blue Flower ◽  
Uridine Diphosphate ◽  
Serine Carboxypeptidase ◽  
Genes Encoding ◽  
Ray Florets

Polyacylated anthocyanins with multiple glycosyl and aromatic acyl groups tend to make flowers display bright and stable blue colours. However, there are few studies on the isolation and functional characterization of genes involved in the polyacylated anthocyanin biosynthesis mechanism, which limits the molecular breeding of truly blue flowers. Senecio cruentus is an important potted ornamental plant, and its blue flowers contain 3′,7-polyacylated delphinidin-type anthocyanins that are not reported in any other plants, suggesting that it harbours abundant gene resources for the molecular breeding of blue flowers. In this study, using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis of blue, carmine and white colours of cineraria cultivars “Venezia” (named VeB, VeC, and VeW, respectively), we found that 3′,7-polyacylated anthocyanin, cinerarin, was the main pigment component that determined the blue colour of ray florets of cineraria. Based on the transcriptome sequencing and differential gene expression (DEG) analysis combined with RT- and qRT-PCR, we found two genes encoding uridine diphosphate glycosyltransferase, named ScUGT1 and ScUGT4; two genes encoding acyl-glucoside-dependent glucosyltransferases which belong to glycoside hydrolase family 1 (GH1), named ScAGGT11 and ScAGGT12; one gene encoding serine carboxypeptidase-like acyltransferase ScSCPL2; and two MYB transcriptional factor genes ScMYB2 and ScMYB4, that were specifically highly expressed in the ray florets of VeB, which indicated that these genes may be involved in cinerarin biosynthesis. The function of ScSCPL2 was analysed by virus-induced gene silencing (VIGS) in cineraria leaves combined with HPLC-MS/MS. ScSCPL2 mainly participated in the 3′ and 7-position acylation of cinerarin. These results will provide new insight into the molecular basis of the polyacylated anthocyanin biosynthesis mechanism in higher plants and are of great significance for blue flower molecular breeding of ornamental plants.

scholarly journals Network Analysis of Transcriptome and LC-MS Reveals a Possible Biosynthesis Pathway of Anthocyanins in Dendrobium officinale

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Zhiyao Ren ◽  
Fangning Qiu ◽  
Yinjie Wang ◽  
Wenxia Yu ◽  
Chenxing Liu ◽  
...  
Keyword(s):  
Active Sites ◽  
Anthocyanin Biosynthesis ◽  
Dendrobium Officinale ◽  
Anthocyanin Synthesis ◽  
Anthocyanin Content ◽  
Serine Carboxypeptidase ◽  
Total Anthocyanin ◽  
Total Anthocyanin Content ◽  
Genes Encoding ◽  
Herbal Plant

Anthocyanins, a group of flavonoids, are widely present in plants and determine the colors of the peels of stems, fruits, and flowers. In this study, we used UHPLC-ESI-MS to identify anthocyanins in the herbal plant Dendrobium officinale, which has been used for centuries in China. The results indicated that the total anthocyanin content in samples from Guangxi was the highest. Seven anthocyanins were identified, and the fragmentation pathways were proposed from D. officinale. Most of the identified anthocyanins were composed of cyanidin and sinapoyl groups. We also carried out that the sinapoyl group had active sites on breast cancer receptors by using Schrödinger. The relative levels of the 7 anthocyanins in the samples from the three locations were determined. Transcriptomic analysis was used to analyze the sinapoyl anthocyanin synthesis-related genes in plants, such as genes encoding UGTs and serine carboxypeptidase. We speculated that sinapoyl anthocyanin biosynthesis was associated with the activities of certain enzymes, including chalcone flavonone isomerase-like, hydroxycinnamoyltransferase 1, UGT-83A1, UGT-88B1 isoform X1, serine carboxypeptidase-like 18 isoform X3, and serine carboxypeptidase-like 18.

scholarly journals Isolating an active and inactive CACTA transposon from lettuce color mutants and characterizing their family

2021 ◽  
Author(s):  
Csanad Gurdon ◽  
Alexander Kozik ◽  
Rong Tao ◽  
Alexander Poulev ◽  
Isabel Armas ◽  
...  
Keyword(s):  
Anthocyanin Biosynthesis ◽  
Mobile Element ◽  
Bioinformatic Analysis ◽  
Relative Expression Level ◽  
Gene Coding ◽  
Genes Encoding ◽  
Key Enzyme ◽  
Natural Derivative ◽  
Dark Green ◽  
Transposon Insertions

Abstract Dietary flavonoids play an important role in human nutrition and health. Flavonoid biosynthesis genes have recently been identified in lettuce (Lactuca sativa); however, few mutants have been characterized. We now report the causative mutations in Green Super Lettuce (GSL), a natural light green mutant derived from red cultivar NAR; and GSL-Dark Green (GSL-DG), an olive-green natural derivative of GSL. GSL harbors CACTA 1 (LsC1), a 3.9-kb active nonautonomous CACTA superfamily transposon inserted in the 5′ untranslated region of anthocyanidin synthase (ANS), a gene coding for a key enzyme in anthocyanin biosynthesis. Both terminal inverted repeats (TIRs) of this transposon were intact, enabling somatic excision of the mobile element, which led to the restoration of ANS expression and the accumulation of red anthocyanins in sectors on otherwise green leaves. GSL-DG harbors CACTA 2 (LsC2), a 1.1-kb truncated copy of LsC1 that lacks one of the TIRs, rendering the transposon inactive. RNA-sequencing and reverse transcription quantitative PCR of NAR, GSL, and GSL-DG indicated the relative expression level of ANS was strongly influenced by the transposon insertions. Analysis of flavonoid content indicated leaf cyanidin levels correlated positively with ANS expression. Bioinformatic analysis of the cv Salinas lettuce reference genome led to the discovery and characterization of an LsC1 transposon family with a putative transposon copy number greater than 1,700. Homologs of tnpA and tnpD, the genes encoding two proteins necessary for activation of transposition of CACTA elements, were also identified in the lettuce genome.

Structure, expression, chromosomal location and product of the gene encoding Adh2 in Petunia.

Genetics ◽  
1993 ◽  
Vol 133 (4) ◽  
pp. 999-1007
Author(s):  
R G Gregerson ◽  
L Cameron ◽  
M McLean ◽  
P Dennis ◽  
J Strommer
Keyword(s):  
Chromosomal Location ◽  
Higher Plants ◽  
Gene Encoding ◽  
Genomic Libraries ◽  
Regulatory Strategy ◽  
Adh1 Gene ◽  
Adh Genes ◽  
Genes Encoding ◽  
Adh Gene ◽  
Polymerase Chain

Abstract In most higher plants the genes encoding alcohol dehydrogenase comprise a small gene family, usually with two members. The Adh1 gene of Petunia has been cloned and analyzed, but a second identifiable gene was not recovered from any of three genomic libraries. We have therefore employed the polymerase chain reaction to obtain the major portion of a second Adh gene. From sequence, mapping and northern data we conclude this gene encodes ADH2, the major anaerobically inducible Adh gene of Petunia. The availability of both Adh1 and Adh2 from Petunia has permitted us to compare their structures and patterns of expression to those of the well-studied Adh genes of maize, of which one is highly expressed developmentally, while both are induced in response to hypoxia. Despite their evolutionary distance, evidenced by deduced amino acid sequence as well as taxonomic classification, the pairs of genes are regulated in strikingly similar ways in maize and Petunia. Our findings suggest a significant biological basis for the regulatory strategy employed by these distant species for differential expression of multiple Adh genes.

Functional characterization of chloroplast-targeted RbgA GTPase in higher plants

2017 ◽  
Vol 95 (4-5) ◽  
pp. 463-479 ◽  
Author(s):  
Young Jeon ◽  
Hee-Kyung Ahn ◽  
Yong Won Kang ◽  
Hyun-Sook Pai
Keyword(s):  
Higher Plants ◽  
Functional Characterization

scholarly journals Higher plants contain homologs of the bacterial celA genes encoding the catalytic subunit of cellulose synthase.

1996 ◽  
Vol 93 (22) ◽  
pp. 12637-12642 ◽  
Author(s):  
J. R. Pear ◽  
Y. Kawagoe ◽  
W. E. Schreckengost ◽  
D. P. Delmer ◽  
D. M. Stalker
Keyword(s):  
Higher Plants ◽  
Cellulose Synthase ◽  
Catalytic Subunit ◽  
Genes Encoding

scholarly journals Characterization of a novel 3-O-α-D-galactosyl-α-L-arabinofuranosidase for the assimilation of gum arabic AGP in Bifidobacterium longum subsp. longum

2021 ◽  
Author(s):  
Yuki Sasaki ◽  
Ayako Horigome ◽  
Toshitaka Odamaki ◽  
Jin-Zhong Xiao ◽  
Akihiro Ishiwata ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Bifidobacterium Longum ◽  
Gum Arabic ◽  
Glycoside Hydrolase Family ◽  
Type Ii ◽  
Cooperative Action ◽  
Genes Encoding ◽  
Key Enzyme ◽  
Degradative Enzymes

Gum arabic arabinogalactan (AG) protein (AGP) is a unique dietary fiber that is degraded and assimilated by only specific strains of Bifidobacterium longum subsp. longum. Here, we identified a novel 3-O-α-d-galactosyl-α-l-arabinofuranosidase (GAfase) from B. longum JCM7052, and classified it into the glycoside hydrolase family 39 (GH39). GAfase released α-d-Galp-(1→3)-l-Ara and β-l-Arap-(1→3)-l-Ara from gum arabic AGP and β-l-Arap-(1→3)-l-Ara from larch AGP, and the α-d-Galp-(1→3)-l-Ara release activity was found to be 594-fold higher than that of β-l-Arap-(1→3)-l-Ara. The GAfase gene was part of a gene cluster that included genes encoding a GH36 α-galactosidase candidate and ABC transporters for the assimilation of the released α-d-Galp-(1→3)-l-Ara in B. longum. Notably, when α-d-Galp-(1→3)-l-Ara was removed from gum arabic AGP, it was assimilated by both B. longum JCM7052 and the non-assimilative B. longum JCM1217, suggesting that the removal of α-d-Galp-(1→3)-l-Ara from gum arabic AGP by GAfase permitted the cooperative action with type-II AG degradative enzymes in B. longum. The present study provides new insight into the mechanism of gum arabic AGP degradation in B. longum. IMPORTANCE Bifidobacteria harbor numerous carbohydrate-active enzymes that degrade several dietary fibers in the gastrointestinal tract. B. longum JCM7052 is known to exhibit the ability to assimilate gum arabic AGP, but the key enzyme involved in the degradation of gum arabic AGP remains unidentified. Here, we cloned and characterized a GH39 3-O-α-d-galactosyl-α-l-arabinofuranosidase (GAfase) from B. longum JCM7052. The enzyme was responsible for the release of α-d-Galp-(1→3)-l-Ara and β-l-Arap-(1→3)-l-Ara from gum arabic AGP. The presence of a gene cluster including the GAfase gene is specifically observed in gum arabic AGP assimilative strains. However, GAfase-carrier strains may affect GAfase-noncarrier strains that express other type-II AG degradative enzymes. These findings provide insights into the bifidogenic effect of gum arabic AGP.

scholarly journals Schistosome tegumental ecto-apyrase (SmATPDase1) degrades exogenous pro-inflammatory and pro-thrombotic nucleotides.

2014 ◽  
Author(s):  
Akram A Da'dara ◽  
Rita Bhardwaj ◽  
Yasser MB Ali ◽  
Patrick Skelly
Keyword(s):  
Thrombus Formation ◽  
Functional Characterization ◽  
Host Cells ◽  
Dependent Manner ◽  
Host Immune Responses ◽  
Genes Encoding ◽  
Molecular Patterns ◽  
Parasitic Worms ◽  
Damage Associated Molecular Patterns

Schistosomes are parasitic worms that can survive in the hostile environment of the human bloodstream where they appear refractory to both immune elimination and thrombus formation. We hypothesize that parasite migration in the bloodstream can stress the vascular endothelium causing this tissue to release chemicals alerting responsive host cells to the stress. Such chemicals are called damage associated molecular patterns (DAMPs) and among the most potent is the proinflammatory mediator, adenosine triphosphate (ATP). Furthermore, the ATP derivative ADP is a pro-thrombotic molecule that acts as a strong activator of platelets. Schistosomes are reported to possess at their host interactive tegumental surface a series of enzymes that could, like their homologs in mammals, degrade extracellular ATP and ADP. These are alkaline phosphatase (SmAP), phosphodiesterase (SmNPP-5) and ATP diphosphohydrolase (SmATPDase1). In this work we employ RNAi to knock down expression of the genes encoding these enzymes in the intravascular life stages of the parasite. We then compare the abilities of these parasites to degrade exogenously added ATP and ADP. . We find that only SmATPDase1-suppressed parasites are significantly impaired in their ability to degrade these nucleotides. Suppression of SmAP or SmNPP-5 does not appreciably affect the worms’ ability to catabolize ATP or ADP. These findings are confirmed by the functional characterization of the enzymatically active, full-length recombinant SmATPDase1 expressed in CHO-S cells. The enzyme is a true apyrase; SmATPDase1 degrades ATP and ADP in a cation dependent manner. Optimal activity is seen at alkaline pH. The Km of SmATPDase1 for ATP is 0.4 ±0.02 mM and for ADP, 0.252 ± 0.02 mM. The results confirm the role of tegumental SmATPDase1 in the degradation of the exogenous pro-inflammatory and pro-thrombotic nucleotides ATP and ADP by live intravascular stages of the parasite. By degrading host inflammatory signals like ATP, and pro-thrombotic signals like ADP, these parasite enzymes may minimize host immune responses, inhibit blood coagulation and promote schistosome survival.)

Sign in / Sign up

Export Citation Format

Share Document