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Article ; Online: COI1 dependent jasmonic acid signalling positively modulates ROS scavenging system in transgenic hairy root culture of tomato.

Kadam, Swapnil B / Barvkar, Vitthal T

2023 Volume 206, Page(s) 108229

Abstract: Reactive oxygen species (ROS) production is a routine event in plants. ROS function as signalling molecules in regulating plant development and defence. However, their accumulation beyond threshold leads to toxicity. Hence, plants are evolved with ... ...

Abstract	Reactive oxygen species (ROS) production is a routine event in plants. ROS function as signalling molecules in regulating plant development and defence. However, their accumulation beyond threshold leads to toxicity. Hence, plants are evolved with specialized ROS scavenging system involving phytohormones (synthesis and signalling), enzymes and metabolites. To understand the role of phytohormone jasmonic acid (JA) signalling in ROS scavenging, tomato coronatine insensitive 1 (SlCOI1), a key gene in JA signalling, was silenced and overexpressed in tomato transgenic hairy roots (HR) under the constitutive promoter. Targeted metabolomics of transgenic HR revealed accumulation of phenolic acids including ferulic acid, coumaric acid, vanillic acid, and flavonoid catechin in SlCOI1 overexpressed line. Moreover, osmolyte amino acids proline, asparagine, and glutamine showed a positive co-relation with transgenic overexpression of SlCOI1. Ascorbic acid-glutathione, a crucial antioxidant system was found to be influenced by COI1-mediated JA signalling. The expression of genes encoding enzymes superoxide dismutase 1, ascorbate peroxidase 1, and dehydroascorbate reductase 2 was found to be down and upregulated in SlCOI1 silenced and overexpressed lines, respectively. Methyl jasmonate and Fusarium oxysporum f.sp. lycopersici crude extract treatment further confirmed the regulatory role of COI1-mediated JA signalling in regulation of enzymatic components involved in ROS scavenging. The COI1-mediated JA signalling could also elevate the expression of RESPIRATORY BURST OXIDASE HOMOLOG-B gene which is involved in ROS wave signal generation. The present study underscores the role of COI1-mediated JA signalling in modulating enzymatic and non-enzymatic components of ROS scavenging system and pathogen associated molecular pattern triggered immunity.
MeSH term(s)	Solanum lycopersicum/genetics ; Reactive Oxygen Species/metabolism ; Oxylipins/metabolism ; Cyclopentanes/metabolism ; Plant Growth Regulators/metabolism ; Gene Expression Regulation, Plant ; Arabidopsis Proteins/genetics
Chemical Substances	jasmonic acid (6RI5N05OWW) ; Reactive Oxygen Species ; Oxylipins ; Cyclopentanes ; Plant Growth Regulators ; Arabidopsis Proteins
Language	English
Publishing date	2023-11-25
Publishing country	France
Document type	Journal Article
ZDB-ID	742978-2
ISSN	1873-2690 ; 0981-9428
ISSN (online)	1873-2690
ISSN	0981-9428
DOI	10.1016/j.plaphy.2023.108229
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Article ; Online: Understanding parasitism in Loranthaceae: Insights from plastome and mitogenome of Helicanthes elastica.

Darshetkar, Ashwini M / Pable, Anupama A / Nadaf, Altafhusain B / Barvkar, Vitthal T

Gene

2023 Volume 861, Page(s) 147238

Abstract: Loranthaceae is the largest family of the order Santalales and includes root and stem hemiparasites. The parasites are known to exhibit reductions in the genomic features as well as relaxed or intensified selection shifts. In this study, we report ... ...

Abstract	Loranthaceae is the largest family of the order Santalales and includes root and stem hemiparasites. The parasites are known to exhibit reductions in the genomic features as well as relaxed or intensified selection shifts. In this study, we report plastome and mitogenome sequence of Helicanthes elastica (subtribe Amyeminae, tribe Lorantheae), an endemic, monotypic genus of Western Ghats, India growing on remarkably diverse host range. The length of plastome sequence was 1,28,805 bp while that of mitogenome was 1,65,273 bp. This is the smallest mitogenome from Loranthaceae reported till date. The plastome of Helicanthes exhibited loss of ndh genes (ψndhB), ψinfA, rps15, rps16, rpl32, trnK-UUU, trnG-UCC, trnV-UAC and trnA-UGC while mitogenome exhibited pseudogenized cox2, nad1 and nad4 genes. The comparative study of Loranthaceae plastomes revealed that the pseudogenization or loss of genes was not specific to any genus or tribe and variation was noted in the number of introns of clpP gene in the family. Several photosynthetic genes have undergone relaxed selection supporting lower photosynthetic rates in parasitic plants while some respiratory genes exhibited intensified selection supporting the idea of host-parasite arm race in Loranthaceae. The plastome gene content was found conserved in root hemiparasites compared to stem hemiparasites. The atp1 gene of mitogenome was chimeric and part of it exhibited similarities with Lamiales members. The phylogenetic analysis based on plastid genes placed Helicanthes sister to the members of subtribe Dendrophthoinae.
MeSH term(s)	Phylogeny ; Loranthaceae ; Rubber ; Genome, Mitochondrial ; Evolution, Molecular
Chemical Substances	Rubber (9006-04-6)
Language	English
Publishing date	2023-02-02
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2023.147238
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Article ; Online: Thidiazuron-mediated callogenesis and biosynthesis of anti-cancerous monoterpene indole alkaloid camptothecin in Nothapodytes nimmoniana (J.Graham) Mabb. callus culture

Kadam, Swapnil B. / Godbole, Rucha C. / Pable, Anupama A. / Singh, Sudhir / Barvkar, Vitthal T.

SAAB South African Journal of Botany. 2023 May, v. 156 p.411-419

2023

Abstract: Nothapodytes nimmoniana is an endangered medicinal plant known for the synthesis of the anti-cancer drug Camptothecin (CPT). Although CPT is considered as a great boon to cancer therapy, its commercial production relies entirely on naturally occurring ... ...

Abstract	Nothapodytes nimmoniana is an endangered medicinal plant known for the synthesis of the anti-cancer drug Camptothecin (CPT). Although CPT is considered as a great boon to cancer therapy, its commercial production relies entirely on naturally occurring plant sources. This resulted in the rampant harvesting of the N. nimmoniana plant population, making the plant endangered. The in vitro system of callus induction and culture faces several challenges, such as recalcitrancy and reduction in CPT content in N. nimmoniana. Here, we report Thidiazuron (TDZ) induced callogenesis and the effect of its different concentrations on the CPT content in the callus. The best explant for callogenesis was found to be hypocotyl, followed by leaf. The TDZ, in combination with 2,4-D, was the most effective among all plant growth regulators combinations tested. The TDZ could induce callus at 1 and 2 mg/L concentrations combined with 0.5 mg/L 2,4-D. The CPT content of the calli maintained on 2 mg/L TDZ+0.5 mg/L 2,4-D was two-fold higher than that of calli on 1 mg/L TDZ+0.5 mg/L 2,4-D. The callus cultures maintained at the selected medium were growing with no signs of organogenesis and necrosis even after multiple sub-cultures. The finding of the experiment emphasizes that TDZ not only induces callogenesis but may also induce CPT biosynthesis in a dose-dependent manner. The results can further be utilized to develop N. nimmoniana callus culture with high CPT content using variable concentrations of TDZ combined with elicitor treatments.
Keywords	2,4-D ; antineoplastic agents ; biosynthesis ; callus ; callus culture ; callus formation ; cancer therapy ; dose response ; elicitors ; hypocotyls ; indole alkaloids ; leaves ; medicinal plants ; monoterpenoids ; necrosis ; organogenesis ; plant growth ; thidiazuron ; Nothapodytes nimmoniana ; Phytohormone ; Camptothecin
Language	English
Dates of publication	2023-05
Size	p. 411-419.
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	2126918-X
ISSN	0254-6299
ISSN	0254-6299
DOI	10.1016/j.sajb.2023.03.022
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Article ; Online: Artificial microRNA mediated silencing of cyclase and aldo–keto reductase genes reveal their involvement in the plumbagin biosynthetic pathway

Vasav, Arati P. / Meshram, Balu G. / Pable, Anupama A. / Barvkar, Vitthal T.

J Plant Res. 2023 Jan., v. 136, no. 1 p.47-62

2023

Abstract: Plumbagin and other naphthoquinone derivatives from the Plumbago zeylanica L. (Plumbaginaceae) are known for their anticancer and other medicinal properties. Previous reports suggest that 3-methyl-1,8-naphthalene-diol is an intermediate of the plumbagin ... ...

Abstract	Plumbagin and other naphthoquinone derivatives from the Plumbago zeylanica L. (Plumbaginaceae) are known for their anticancer and other medicinal properties. Previous reports suggest that 3-methyl-1,8-naphthalene-diol is an intermediate of the plumbagin biosynthetic pathway and is synthesized from hexaketide backbone; a reaction catalyzed by type III polyketide synthase (PKS) along with certain accessory enzymes. Our earlier transcriptomic and metabolomic studies suggest that along with PKS, putative cyclase and aldo–keto reductase might be involved in the formation of 3-methyl-1,8-naphthalene-diol. The present study probed young leaf transcriptome and identified cyclase and aldo–keto reductase like transcripts that might be involved in the intramolecular aldol condensation of hexaketide intermediate and decarboxylation, carbonyl reduction and hydroxyl elimination of keto or enol forms of hexaketide intermediates respectively. Moreover, sequence alignment of identified cyclase1 possesses signature β–α–β–β–α–α–β topology, which belongs to the dimeric α + β barrel (DABB) protein family and is involved in the C2–C11 and C4–C9 intramolecular aldol condensation of hexaketide intermediates. Along with cyclase1, we further identified and characterized P. zeylanica specific aldo–keto reductase1 (AKR1) which is a novel member of the aldo–keto reductase (AKR) multi-gene family that possesses the conserved Asp60, Tyr65, Lys91, and His132 residues and is proposed to be involved in the C1 decarboxylation, C3 carbonyl reduction and C7 hydroxyl elimination of keto or enol form of hexaketide intermediate to form 3-methyl-1,8-naphthalene-diol. Further, the functional characterization using the artificial microRNA mediated transient silencing approach confirmed the involvement of cyclase1 and AKR1 in the plumbagin biosynthetic pathway. This is the first study reporting the identification and functional characterization of cyclase1 and AKR1 genes involved in the plumbagin biosynthetic pathway and general plant polyketide biosynthesis.
Keywords	Plumbago zeylanica ; aldo-keto reductases ; biochemical pathways ; biosynthesis ; condensation reactions ; decarboxylation ; enols ; leaves ; metabolomics ; microRNA ; multigene family ; plumbagin ; polyketide synthases ; polyketides ; sequence alignment ; topology ; transcriptome ; transcriptomics
Language	English
Dates of publication	2023-01
Size	p. 47-62.
Publishing place	Springer Nature Singapore
Document type	Article ; Online
ZDB-ID	2077362-6
ISSN	1618-0860 ; 0918-9440
ISSN (online)	1618-0860
ISSN	0918-9440
DOI	10.1007/s10265-022-01415-7
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Article ; Online: Understanding parasitism in Loranthaceae: Insights from plastome and mitogenome of Helicanthes elastica

Darshetkar, Ashwini M. / Pable, Anupama A. / Nadaf, Altafhusain B. / Barvkar, Vitthal T.

Gene. 2023 Apr., v. 861 p.147238-

2023

Abstract	Loranthaceae is the largest family of the order Santalales and includes root and stem hemiparasites. The parasites are known to exhibit reductions in the genomic features as well as relaxed or intensified selection shifts. In this study, we report plastome and mitogenome sequence of Helicanthes elastica (subtribe Amyeminae, tribe Lorantheae), an endemic, monotypic genus of Western Ghats, India growing on remarkably diverse host range. The length of plastome sequence was 1,28,805 bp while that of mitogenome was 1,65,273 bp. This is the smallest mitogenome from Loranthaceae reported till date. The plastome of Helicanthes exhibited loss of ndh genes (ψndhB), ψinfA, rps15, rps16, rpl32, trnK-UUU, trnG-UCC, trnV-UAC and trnA-UGC while mitogenome exhibited pseudogenized cox2, nad1 and nad4 genes. The comparative study of Loranthaceae plastomes revealed that the pseudogenization or loss of genes was not specific to any genus or tribe and variation was noted in the number of introns of clpP gene in the family. Several photosynthetic genes have undergone relaxed selection supporting lower photosynthetic rates in parasitic plants while some respiratory genes exhibited intensified selection supporting the idea of host-parasite arm race in Loranthaceae. The plastome gene content was found conserved in root hemiparasites compared to stem hemiparasites. The atp1 gene of mitogenome was chimeric and part of it exhibited similarities with Lamiales members. The phylogenetic analysis based on plastid genes placed Helicanthes sister to the members of subtribe Dendrophthoinae.
Keywords	Lamiales ; Loranthaceae ; comparative study ; genomics ; host range ; introns ; mitochondrial genome ; parasitism ; photosynthesis ; phylogeny ; India ; Biorganellar genomes ; Evolution ; Intensified selection ; Relaxed selection ; Santalales
Language	English
Dates of publication	2023-04
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2023.147238
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Article ; Online: Trichoderma virens exerts herbicidal effect on Arabidopsis thaliana via modulation of amino acid metabolism

Bansal, Ravindra / Sahoo, Sripati Abhiram / Barvkar, Vitthal Tanaji / Srivastava, Ashish Kumar / Mukherjee, Prasun Kumar

Plant Science. 2023 July, v. 332 p.111702-

2023

Abstract: Trichoderma virens is a plant beneficial fungus well-known for its biocontrol, herbicidal and growth promotion activity. Earlier, we identified HAS (HA-synthase, a terpene cyclase) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) to be involved in ... ...

Abstract	Trichoderma virens is a plant beneficial fungus well-known for its biocontrol, herbicidal and growth promotion activity. Earlier, we identified HAS (HA-synthase, a terpene cyclase) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) to be involved in the production of multiple non-volatiles and non-volatile+volatile metabolites, respectively. The present study delineates the function of HAS and GAPDH in regulating herbicidal activity, using the model plant Arabidopsis thaliana. Under axenic conditions, rosette-biomass of seedlings co-cultivated with ΔHAS (HASᴿ) and ΔGAPDH (GAPDHᴿ) was higher than WT-Trichoderma (WTᴿ) as well as non-colonized control (NoTᴿ), even though the root colonization ability was reduced. However, HASᴿ biomass was still higher than those of GAPDHᴿ, indicating that blocking volatiles will not provide any additional contribution over non-volatile metabolites for Trichoderma-induced herbicidal activity. LC-MS analysis revealed that loss of herbicidal activity of ΔHAS/ΔGAPDH was associated with an increase in the levels of amino acids, which coincided with reduced expression levels of amino-acid catabolism and anabolism related genes in HASᴿ/GAPDHᴿ. RNAi-mediated suppression of an oxidoreductase gene, VDN5, specifically prevented viridin-to-viridiol conversion. Additionally, vdn5 mimics ΔHAS, in terms of amino-acid metabolism gene expression and partially abolishes the herbicidal property of WT-Trichoderma. Thus, the study provides mechanistic frame-work for better utilization of Trichoderma virens for biocontrol purposes, balancing between plant growth promotion and herbicidal activity.
Keywords	Arabidopsis thaliana ; Trichoderma virens ; amino acid metabolism ; amino acids ; anabolism ; biological control ; biomass ; catabolism ; colonizing ability ; fungi ; gene expression ; genes ; glyceraldehyde-3-phosphate dehydrogenase ; growth promotion ; herbicidal properties ; metabolites ; plant growth ; Biocontrol ; Plant growth promotion ; Secondary metabolites ; Viridin ; Viridiol ; Volatiles
Language	English
Dates of publication	2023-07
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	742010-9
ISSN	1873-2259 ; 0168-9452
ISSN (online)	1873-2259
ISSN	0168-9452
DOI	10.1016/j.plantsci.2023.111702
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Article ; Online: Phylogenomics of transcriptionally active AP2/ERF and bHLH transcription factors and study of their promoter regions in

Godbole, Rucha C / Kadam, Swapnil B / Pable, Anupama A / Singh, Sudhir / Barvkar, Vitthal T

Genome

2023 Volume 66, Issue 9, Page(s) 235–250

Abstract: Nothapodytes ... ...

Abstract	Nothapodytes nimmoniana
MeSH term(s)	Humans ; Basic Helix-Loop-Helix Transcription Factors/genetics ; Basic Helix-Loop-Helix Transcription Factors/metabolism ; Phylogeny ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Alkaloids ; Plants, Medicinal/genetics ; Promoter Regions, Genetic ; Plant Proteins/genetics ; Plant Proteins/metabolism ; Gene Expression Regulation, Plant ; Repressor Proteins/genetics ; Repressor Proteins/metabolism
Chemical Substances	Basic Helix-Loop-Helix Transcription Factors ; Transcription Factors ; Alkaloids ; Plant Proteins ; ERF protein, human ; Repressor Proteins
Language	English
Publishing date	2023-05-10
Publishing country	Canada
Document type	Journal Article
ZDB-ID	639031-6
ISSN	1480-3321 ; 0831-2796
ISSN (online)	1480-3321
ISSN	0831-2796
DOI	10.1139/gen-2023-0009
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Article ; Online: Artificial microRNA mediated silencing of cyclase and aldo-keto reductase genes reveal their involvement in the plumbagin biosynthetic pathway.

Vasav, Arati P / Meshram, Balu G / Pable, Anupama A / Barvkar, Vitthal T

Journal of plant research

2022

Abstract	Plumbagin and other naphthoquinone derivatives from the Plumbago zeylanica L. (Plumbaginaceae) are known for their anticancer and other medicinal properties. Previous reports suggest that 3-methyl-1,8-naphthalene-diol is an intermediate of the plumbagin biosynthetic pathway and is synthesized from hexaketide backbone; a reaction catalyzed by type III polyketide synthase (PKS) along with certain accessory enzymes. Our earlier transcriptomic and metabolomic studies suggest that along with PKS, putative cyclase and aldo-keto reductase might be involved in the formation of 3-methyl-1,8-naphthalene-diol. The present study probed young leaf transcriptome and identified cyclase and aldo-keto reductase like transcripts that might be involved in the intramolecular aldol condensation of hexaketide intermediate and decarboxylation, carbonyl reduction and hydroxyl elimination of keto or enol forms of hexaketide intermediates respectively. Moreover, sequence alignment of identified cyclase1 possesses signature β-α-β-β-α-α-β topology, which belongs to the dimeric α + β barrel (DABB) protein family and is involved in the C2-C11 and C4-C9 intramolecular aldol condensation of hexaketide intermediates. Along with cyclase1, we further identified and characterized P. zeylanica specific aldo-keto reductase1 (AKR1) which is a novel member of the aldo-keto reductase (AKR) multi-gene family that possesses the conserved Asp60, Tyr65, Lys91, and His132 residues and is proposed to be involved in the C1 decarboxylation, C3 carbonyl reduction and C7 hydroxyl elimination of keto or enol form of hexaketide intermediate to form 3-methyl-1,8-naphthalene-diol. Further, the functional characterization using the artificial microRNA mediated transient silencing approach confirmed the involvement of cyclase1 and AKR1 in the plumbagin biosynthetic pathway. This is the first study reporting the identification and functional characterization of cyclase1 and AKR1 genes involved in the plumbagin biosynthetic pathway and general plant polyketide biosynthesis.
Language	English
Publishing date	2022-10-13
Publishing country	Japan
Document type	Journal Article
ZDB-ID	2077362-6
ISSN	1618-0860 ; 0918-9440
ISSN (online)	1618-0860
ISSN	0918-9440
DOI	10.1007/s10265-022-01415-7
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Article ; Online: Evidence of polyamines mediated 2-acetyl-1-pyrroline biosynthesis in aromatic rice rhizospheric fungal species Aspergillus niger.

Zehra, Andleeb / Dhondge, Harshal V / Barvkar, Vitthal T / Singh, Sanjay K / Nadaf, Altafhusain B

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology

2023 Volume 54, Issue 4, Page(s) 3073–3083

Abstract: Rhizosphere soil of aromatic rice inhabits different fungal species that produce many bioactive metabolites including 2-acetyl-1-pyrroline (2AP). The mechanism for the biosynthesis of 2AP in the fungal system is still elusive. Hence, the present study ... ...

Abstract	Rhizosphere soil of aromatic rice inhabits different fungal species that produce many bioactive metabolites including 2-acetyl-1-pyrroline (2AP). The mechanism for the biosynthesis of 2AP in the fungal system is still elusive. Hence, the present study investigates the role of possible nitrogen (N) precursors such as some amino acids and polyamines as well as the enzymes involved in 2AP synthesis in the fungal species isolated from the rhizosphere of aromatic rice varieties. Three fungal isolates were found to synthesize 2AP (0.32-1.07 ppm) and maximum 2AP was synthesized by Aspergillus niger (1.07 ppm) isolated from rhizosphere of Dehradun Basmati (DB). To determine the N source for 2AP synthesis, various N sources such as proline, glutamate, ornithine putrescine, spermine, and spermidine were used in place of putrescine in the synthetic medium (Syn18). The results showed that maximum 2AP synthesis was found with putrescine (1.07 ppm) followed by spermidine (0.89 ppm) and spermine (0.84 ppm). Further, LC-QTOF-MS analysis revealed the mobilization of spermine and spermidine into the putrescine, indicating that putrescine is the key N source for 2AP synthesis. Moreover, higher enzyme activity of DAO, PAO, and ODC as well as higher content of methylglyoxal metabolite in the A. niger NFCCI 5060 as compared to A. niger NFCCI 4064 (control) suggests the prominent role of these enzymes in the synthesis of 2AP. In conclusion, this study showed evidence of the polyamines mediated 2AP biosynthesis in A. niger NFCCI 5060.
MeSH term(s)	Polyamines/metabolism ; Spermidine/metabolism ; Putrescine/metabolism ; Spermine/metabolism ; Aspergillus niger/genetics ; Aspergillus niger/metabolism ; Oryza/metabolism ; Ornithine Decarboxylase/metabolism
Chemical Substances	Polyamines ; Spermidine (U87FK77H25) ; Putrescine (V10TVZ52E4) ; Spermine (2FZ7Y3VOQX) ; 2-acetyl-1-pyrroline (IGC0W6LY94) ; Ornithine Decarboxylase (EC 4.1.1.17)
Language	English
Publishing date	2023-09-13
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	2017175-4
ISSN	1678-4405 ; 1517-8382
ISSN (online)	1678-4405
ISSN	1517-8382
DOI	10.1007/s42770-023-01124-w
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Article ; Online: Genome sequencing and protein modeling unraveled the 2AP biosynthesis in Bacillus cereus DB25.

Dhondge, Harshal V / Barvkar, Vitthal T / Dastager, Syed G / Dharne, Mahesh S / Rajput, Vinay / Pable, Anupama A / Henry, Robert J / Nadaf, Altafhusain B

International journal of food microbiology

2024 Volume 413, Page(s) 110600

Abstract: 2-Acetyl-1-pyrroline (2AP) is an important and major flavor aroma compound responsible for the fragrance of basmati rice, cheese, wine, and several other food products. Biosynthesis of 2AP in aromatic rice and a few other plant species is associated with ...

Abstract	2-Acetyl-1-pyrroline (2AP) is an important and major flavor aroma compound responsible for the fragrance of basmati rice, cheese, wine, and several other food products. Biosynthesis of 2AP in aromatic rice and a few other plant species is associated with a recessive Betaine aldehyde dehydrogenase 2 (BADH2) gene. However, the literature is scant on the relationship between the functional BADH2 gene and 2AP biosynthesis in prokaryotic systems. Therefore, in the present study, we aimed to explore the functionality of the BADH2 gene for 2AP biosynthesis in 2AP synthesizing rice rhizobacterial isolate Bacillus cereus DB25 isolated from the rhizosphere of basmati rice (Oryza sativa L.). Full-length BcBADH2 sequence was obtained through whole genome sequencing (WGS) and further confirmed through traditional PCR and Sanger sequencing. Then the functionality of the BcBADH2 gene was evaluated in-silico through bioinformatics analysis and protein docking studies and further experimentally validated through enzyme assay. The sequencing and bioinformatics analysis results revealed a full-length 1485 bp BcBADH2 coding sequence without any deletion or premature stop codons. Full-length BcBADH2 was found to encode a fully functional protein of 54.08 kDa with pI of 5.22 and showed the presence of the conserved amino acids responsible for enzyme activity. The docking studies confirmed a good affinity between the protein and its substrate whereas the presence of BcBADH2 enzyme activity confirmed the functionality of BADH2 enzyme in B. cereus DB25. In conclusion, the findings of the present study suggest that B. cereus DB25 is able to synthesize 2AP despite a functional BADH2 gene and there may be a different molecular mechanism responsible for 2AP biosynthesis in bacterial systems, unlike that found in aromatic rice and other eukaryotic plant species.
MeSH term(s)	Bacillus cereus/genetics ; Bacillus cereus/metabolism ; Base Sequence ; Odorants/analysis ; Plant Proteins/metabolism ; Oryza ; Pyrroles/metabolism
Chemical Substances	Plant Proteins ; Pyrroles
Language	English
Publishing date	2024-01-26
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	87122-9
ISSN	1879-3460 ; 0168-1605
ISSN (online)	1879-3460
ISSN	0168-1605
DOI	10.1016/j.ijfoodmicro.2024.110600
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