Abstract
Polyamines and their derivatives, such as phenolamides, play essential roles in plant defence systems, especially under biotic stress conditions (Gerlin et al. 2021). Anthracnose, caused by Colletotrichum gloeosporioides, is a major postharvest disease affecting mango (Mangifera indica L.), leading to significant economic losses during storage and transportation. Sustainable approaches that leverage mango's intrinsic defence mechanisms offer promising alternatives for managing postharvest diseases. This study identifies p-coumaroyl-feruloyl-putrescine (CFP), a phenolamide derived from polyamines, as a critical metabolite in mango's defence against anthracnose, and investigates its biosynthesis, regulation, and antifungal efficacy. To identify key metabolites involved in mango's response to anthracnose, we conducted a time-course metabolomic analysis of infected (T1d to T3d, 1–3 days post-inoculation) and control (CK0d to CK3d) fruit samples, with each sample comprising three biological replicates. Using UPLC–MS, we identified 175 distinct alkaloids, including phenolamides (Table S1). Metabolite values were normalised using Z-scores before performing orthogonal partial least squares discriminant analysis (OPLS-DA). The OPLS-DA model revealed distinct metabolic profiles between infected and control samples, highlighting infection-driven metabolic reprogramming (Figure 1a). Among the metabolites, CFP emerged as the most significant, exhibiting the highest score and a pronounced accumulation during the early stages of infection (Figure 1a–c; Tables S2 and S3). These findings suggest that CFP plays a central role as a rapidly induced defence metabolite during the initial response to anthracnose. To elucidate the biosynthetic pathways underlying CFP production, we conducted a transcriptomic analysis to identify genes associated with CFP production (Tables S4 and S5). k-means clustering was performed using the Euclidean distance metric. Metabolites and genes were grouped into five distinct clusters (Figure 1d). Cluster 3, containing CFP and seven other metabolites, displayed a sharp increase at 1 and 2 dpi, followed by a decline at 3 dpi, reflecting an early and transient defence response. The genes in cluster 3 showed expression profiles closely mirroring the temporal patterns observed in metabolomic cluster 3, confirming a coordinated activation of metabolic and transcriptional defences. In this cluster, a series of genes involved in phenylpropanoid and polyamine pathways were identified. Co-expression analysis further linked these genes to CFP biosynthesis (Figure 1e; Table S6), suggesting that phenylpropanoid and polyamine pathway genes are temporally and spatially regulated to regulate CFP biosynthesis. Then we proposed the biosynthetic pathways leading to CFP synthesis by integrating both the phenylpropanoid and polyamine pathways (Figure 1f; Table S7). Enzymes in the phenylpropanoid pathway, including phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), and caffeic acid O-methyltransferase (OMT), were significantly upregulated during infection. These enzymes synthesise the acyl donors p-coumaroyl-CoA and feruloyl-CoA (Wang et al. 2023). Similarly, arginine decarboxylase (ADC), a critical enzyme in the polyamine pathway, showed elevated expression, producing putrescine as the amine donor for CFP synthesis. The final steps of CFP biosynthesis involve the conjugation of p-coumaroyl-CoA and feruloyl-CoA with putrescine, catalysed by acyltransferase (AT) and berberine bridge enzyme-like (BBL) proteins (Bai et al. 2022). To identify regulatory elements involved in CFP biosynthesis, we focused on MYB14 and MYB15, two Jasmonic acid (JA) responsive transcription factors (He et al. 2020). These regulators showed strong co-expression with CFP biosynthetic genes (Figure 1e), indicating their central role in activating CFP production. Functional validation of CFP biosynthetic genes was performed using transient overexpression in Nicotiana benthamiana leaves (Figure 1g; Table S8). Overexpression of individual biosynthetic genes did not significantly increase CFP accumulation. However, the expression of MYB14 or MYB15 led to substantial production of CFP and its precursor, feruloyl-putrescine (FP), reinforcing the regulatory role of these transcription factors in orchestrating the phenylpropanoid and polyamine pathways. Additionally, exogenous JA treatments significantly enhanced the expression of MYB14, MYB15, and CFP-related genes, further supporting the role of JA as a key hormonal signal driving phenolamide-mediated defence. In contrast, salicylic acid (SA) treatments had minimal effects on CFP biosynthetic gene expression (Figure 1h). To assess the antifungal efficacy of CFP, its precursors, putrescine and FP, were selected due to the unavailability of commercially standardised CFP compounds. We conducted in vitro and in vivo assays (Figure S1). In vitro antifungal activity was evaluated by incorporating putrescine and FP into potato dextrose agar (PDA) medium and measured fungal colony growth to assess inhibitory effects. For in vivo assays, mango and apple fruits were inoculated with C. gloeosporioides conidia, followed by the application of putrescine or FP to the infection site. In vitro tests revealed that neither putrescine nor FP inhibited fungal growth on PDA plates, indicating limited direct antifungal activity under these conditions. However, in vivo assays demonstrated significant reductions in anthracnose lesion sizes on mango and apple fruits treated with putrescine or FP. FP exhibited greater protective effects than putrescine, reducing lesion sizes by approximately 30% compared to untreated controls. This context-specific activity suggests that CFP and its precursors function by modulating host defences and fortifying structural barriers, rather than directly inhibiting fungal growth (Zeiss et al. 2021). In conclusion, CFP plays a pivotal role in mango's defence against anthracnose, synthesised through the coordinated action of phenylpropanoid and polyamine pathways and regulated by MYB14 and MYB15 under JA signalling (Figure 1i). B.A. and B.Z. designed the project. B.Z., L.H., Q.X., and Q.W. performed the experiments. B.Z., L.H. and Q.W. analysed the data. B.A., B.Z., Q.W. and H.L. wrote the manuscript. This research was funded by the Central Public-interest Scientific Institution Basal Research Fund for CATAS (1630032022007); National Natural Science Foundation of China (32160594, 32260042). The data that support the findings of this study are available in the Supporting Information of this article. Figure S1. Tables S1–S8. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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Claims 19
Claim · model-written summary
UPLC–MS identified 175 alkaloids including phenolamides.
- Taxon
- mango
_from_India.jpg?width=60)
Mangifera indicat0 - Outcome
- 175 distinct alkaloidse0
- Value
- 175 distinct alkaloidse0
Not reportedGeneOrgan / stageInteractorTreatmentComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_from_India.jpg?width=500)
Verbatim
e0“Using UPLC–MS, we identified 175 distinct alkaloids, including phenolamides”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Infected and control mango fruit had distinct metabolic profiles.
- Taxon
- mangoMangifera indicat0
- Treatment
- infectede0
- Outcome
- distinct metabolic profilese0
- Direction
- distincte0
- Comparator
- control samplese0
Not reportedGeneOrgan / stageInteractorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The OPLS-DA model revealed distinct metabolic profiles between infected and control samples”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
CFP accumulated prominently during early infection.
- Taxon
- mangoMangifera indicat0
- Treatment
- infectione0
- Outcome
- pronounced accumulatione0
- Direction
- accumulatione0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“CFP emerged as the most significant, exhibiting the highest score and a pronounced accumulation during the early stages of infection”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Metabolites and genes formed five distinct clusters.
- Taxon
- mangoMangifera indicat0
- Outcome
- five distinct clusterse0
- Value
- five distinct clusterse0
Not reportedGeneOrgan / stageInteractorTreatmentComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Metabolites and genes were grouped into five distinct clusters”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Cluster 3 metabolites rose on days 1 and 2 and declined on day 3.
- Taxon
- mangoMangifera indicat0
- Outcome
- CFP and seven other metabolitese0
- Direction
- sharp increasee0
- Value
- 1 and 2 dpie0
Not reportedGeneOrgan / stageInteractorTreatmentComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Cluster 3, containing CFP and seven other metabolites, displayed a sharp increase at 1 and 2 dpi, followed by a decline at 3 dpi”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Genes in cluster 3 mirrored the metabolite time pattern.
- Taxon
- mangoMangifera indicat0
- Gene
- genes in cluster 3e0
- Outcome
- expression profilese0
- Direction
- closely mirroringe0
Not reportedOrgan / stageInteractorTreatmentComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The genes in cluster 3 showed expression profiles closely mirroring the temporal patterns observed in metabolomic cluster 3”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Phenylpropanoid and polyamine pathway genes were identified in cluster 3.
- Taxon
- mangoMangifera indicat0
- Gene
- genes involved in phenylpropanoid and polyamine pathwayse0
- Outcome
- genes involved in phenylpropanoid and polyamine pathwayse0
Not reportedOrgan / stageInteractorTreatmentComparatorDirectionValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In this cluster, a series of genes involved in phenylpropanoid and polyamine pathways were identified”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Co-expression linked pathway genes with CFP biosynthesis.
- Taxon
- mangoMangifera indicat0
- Gene
- these genese0
- Outcome
- CFP biosynthesise0
- Direction
- linkede0
Not reportedOrgan / stageInteractorTreatmentComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Co-expression analysis further linked these genes to CFP biosynthesis”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
PAL, C4H, and 4CL expression rose significantly during infection.
- Taxon
- mangoMangifera indicat0
- Treatment
- infectione0
- Gene
- phenylalanine ammonia-lyase (PAL)e0
- Outcome
- significantly upregulatede0
- Direction
- upregulatede0
Not reportedOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Enzymes in the phenylpropanoid pathway, including phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate-CoA ligase (4CL), and caffeic acid O-methyltransferase (OMT), were significantly upregulated during infection”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
OMT expression rose significantly during infection.
- Taxon
- mangoMangifera indicat0
- Treatment
- infectione0
- Gene
- caffeic acid O-methyltransferase (OMT)e0
- Outcome
- significantly upregulatede0
- Direction
- upregulatede0
Not reportedOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“caffeic acid O-methyltransferase (OMT), were significantly upregulated during infection”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
ADC expression was elevated.
- Taxon
- mangoMangifera indicat0
- Gene
- arginine decarboxylase (ADC)e0
- Outcome
- elevated expressione0
- Direction
- elevatede0
Not reportedOrgan / stageInteractorTreatmentComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“arginine decarboxylase (ADC), a critical enzyme in the polyamine pathway, showed elevated expression”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
MYB14 and MYB15 co-expressed with CFP biosynthetic genes.
- Taxon
- mangoMangifera indicat0
- Gene
- CFP biosynthetic genese0
- Outcome
- strong co-expressione0
- Direction
- strong co-expressione0
Not reportedOrgan / stageInteractorTreatmentComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“These regulators showed strong co-expression with CFP biosynthetic genes”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Individual biosynthetic gene overexpression did not significantly increase CFP accumulation.
- Taxon
- mangoMangifera indicat0
- Treatment
- Overexpression of individual biosynthetic genese0
- Outcome
- CFP accumulatione0
- Direction
- did not significantly increasee0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Overexpression of individual biosynthetic genes did not significantly increase CFP accumulation”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
MYB14 or MYB15 expression produced CFP and feruloyl-putrescine.
- Taxon
- mangoMangifera indicat0
- Treatment
- expression of MYB14 or MYB15e0
- Outcome
- production of CFP and its precursor, feruloyl-putrescine (FP)e0
- Direction
- substantial productione0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the expression of MYB14 or MYB15 led to substantial production of CFP and its precursor, feruloyl-putrescine (FP)”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Exogenous JA enhanced expression of MYB14, MYB15 and CFP-related genes.
- Taxon
- mangoMangifera indicat0
- Treatment
- exogenous JA treatmentse0
- Outcome
- expression of MYB14, MYB15, and CFP-related genese0
- Direction
- enhancede0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“exogenous JA treatments significantly enhanced the expression of MYB14, MYB15, and CFP-related genes”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Salicylic acid treatment had minimal effects on CFP biosynthetic gene expression.
- Taxon
- mangoMangifera indicat0
- Treatment
- salicylic acid (SA) treatmentse0
- Outcome
- CFP biosynthetic gene expressione0
- Direction
- minimal effectse0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“salicylic acid (SA) treatments had minimal effects on CFP biosynthetic gene expression”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Neither putrescine nor FP inhibited fungal growth in vitro.
- Taxon
- mangoMangifera indicat0
- Treatment
- putrescinee0
- Outcome
- fungal growthe0
- Direction
- neither putrescine nor FP inhibitede0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“neither putrescine nor FP inhibited fungal growth on PDA plates”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
Putrescine or FP significantly reduced anthracnose lesions in mango and apple fruits.
- Taxon
- mangoMangifera indicat0
- Treatment
- putrescine or FPe0
- Outcome
- anthracnose lesion sizese0
- Direction
- reductionse0
Not reportedGeneOrgan / stageInteractorComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“significant reductions in anthracnose lesion sizes on mango and apple fruits treated with putrescine or FP”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”
Claim · model-written summary
FP reduced lesion size approximately 30% compared with untreated controls, more than putrescine.
- Taxon
- mangoMangifera indicat0
- Treatment
- FPe0
- Outcome
- lesion sizese0
- Direction
- reducinge0
- Value
- approximately 30%e0
- Comparator
- untreated controlse0
Not reportedGeneOrgan / stageInteractor
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“FP exhibited greater protective effects than putrescine, reducing lesion sizes by approximately 30% compared to untreated controls”
t0“Characterisation of Polyamines and Their Biosynthetic Pathways Contributing to Postharvest Anthracnose Resistance in Mango ( Mangifera indica L.)”