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Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Functional analysis of the CCoAOMT gene in Populus deltoides for enhancing tolerance to Alternaria burnsii

Ankit Yadav A , Amar Sakure A * , Sushil Kumar https://orcid.org/0000-0003-3767-8846 A , Akarsh Parihar B , Dimple Gor A , Vidyut Balar B and Pratibha B
+ Author Affiliations
- Author Affiliations

A Department of Agriculture Biotechnology, Anand Agricultural University, Anand, India. Email: sushil254386@yahoo.com

B Department of Genetics and Plant Breeding, Anand Agricultural University, Anand, India.

* Correspondence to: sakure_amar455@gmail.com

Handling Editor: Inzamam Haq

Functional Plant Biology 52, FP25064 https://doi.org/10.1071/FP25064
Submitted: 21 February 2025  Accepted: 23 April 2025  Published: 8 May 2025

© 2025 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Alternaria blight (Alternaria burnsii) causes significant economic losses due to defoliation, reduced yields, and poor-quality produce in various crops. Consequently, effective strategies for managing this disease are critical. In this study, the caffeoyl-CoA O-methyltransferase (PdCCoAOMT) gene, which plays a key role in lignin biosynthesis and plant defense, was isolated from Populus deltoides and investigated for its potential to enhance resistance against A. burnsii, the causal agent of blight of various crop species. The PdCCoAOMT gene (741 bp) was cloned, characterised, and expressed in the model plant Nicotiana tabacum via Agrobacterium-mediated transformation. Sequencing of the amplicon followed by BLAST analysis revealed 100% query coverage and 98.52% identity of CCoAOMT with the Populus tomentosa and Populus trichocarpa mRNA. Histochemical GUS staining of the putative transformed leaves displayed a distinct blue colour, predominantly in the veins. Gene expression analysis via real time quantitative PCR of 11 T1 plants showed the highest expression in T1-6 plant. Overexpression of PdCCoAOMT gene showed a positive correlation with lignin deposition in the transformed plants compared to the control plants. A detached leaf assay for A. burnsii resistance demonstrated a significant negative correlation between lignin deposition and disease severity, suggesting that higher lignin accumulation in the leaf was associated with reduced disease symptoms. This highlights the effectiveness of the gene in mitigating the disease in the transformed tobacco plants. These findings suggest that PdCCoAOMT could be a valuable tool in developing crop varieties resistant to Alternaria blight, providing a promising strategy to combat this economically devastating pathogen.

Keywords: Alternaria burnsii, blight, CCoAOMT, disease resistance, gene cloning, gene expression, lignin, Populus.

References

Ahmed SS, Rifat ZT, Lohia R, Campbell AJ, Dunker AK, Rahman MS, Iqbal S (2022) Characterization of intrinsically disordered regions in proteins informed by human genetic diversity. PLOS Computational Biology 18(3), e1009911.
| Crossref | Google Scholar | PubMed |

Busam G, Junghanns KT, Kneusel RE, Kassemeyer HH, Matern U (1997) Characterization and expression of caffeoyl-coenzyme A 3-O-methyltransferase proposed for the induced resistance response of Vitis vinifera L. Plant Physiology 115(3), 1039-1048.
| Crossref | Google Scholar | PubMed |

Chen C, Meyermans H, Burggraeve B, De Rycke RM, Inoue K, De Vleesschauwer V, Steenackers M, Van Montagu MC, Engler GJ, Boerjan WA (2000) Cell-specific and conditional expression of caffeoyl-coenzyme A-3-O-methyltransferase in poplar. Plant Physiology 123(3), 853-868.
| Crossref | Google Scholar | PubMed |

Choubey VK, Sakure AA, Kumar S, Vaja MB, Mistry JG, Patel DA (2023) Proteomics profiling and in silico analysis of peptides identified during Fusarium oxysporum infection in castor (Ricinus communis). Phytochemistry 213, 113776.
| Crossref | Google Scholar | PubMed |

Ferrer J-L, Zubieta C, Dixon RA, Noel JP (2005) Crystal structures of alfalfa caffeoyl coenzyme A 3-O-methyltransferase. Plant Physiology 137(3), 1009-1017.
| Crossref | Google Scholar | PubMed |

Gargouri M, Selmi S, Fardeau ML, Sadfi-Zouaoui N (2022) Diversity and ecological roles of saprophytic fungi in soil ecosystems: focus on Alternaria species. Journal of Fungi 8(3), 250.
| Google Scholar |

Ge C, Wang Y-G, Lu S, Zhao XY, Hou B-K, Balint-Kurti PJ, Wang G-F (2021) Multi-omics analyses reveal the regulatory network and the function of ZmUGTs in maize defense response. Frontiers in Plant Science 12, 738261.
| Crossref | Google Scholar | PubMed |

Giordano D, Provenzano S, Ferrandino A, Vitali M, Pagliarani C, Roman F, Cardinale F, Castellarin SD, Schubert A (2016) Characterization of a multifunctional caffeoyl-CoA O-methyltransferase activated in grape berries upon drought stress. Plant Physiology and Biochemistry 101, 23-32.
| Crossref | Google Scholar | PubMed |

Gregersen PL, Christensen AB, Sommer-Knudsen J, Collinge DB (1994) A putative O-methyltransferase from barley is induced by fungal pathogens and UV light. Plant Molecular Biology 26(6), 1797-1806.
| Crossref | Google Scholar | PubMed |

Hegde N, Joshi S, Soni N, Kushalappa AC (2021) The caffeoyl-CoA O-methyltransferase gene SNP replacement in Russet Burbank potato variety enhances late blight resistance through cell wall reinforcement. Plant Cell Reports 40(1), 237-254.
| Crossref | Google Scholar | PubMed |

Horsch RB, Fry JE, Hoffmann NL, Wallroth M, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227(4691), 1229-1231.
| Crossref | Google Scholar |

Jenny P, Sakure AA, Yadav A, Kumar S (2024) Molecular cloning and characterisation of root-specific SlREO promoter of the Indian tomato (Solanum lycopersicum L.) cultivar. Functional Plant Biology 51, FP24063.
| Crossref | Google Scholar |

Kelley LA, Mezulis S, Yates CM, Wass MN, Sternberg MJE (2015) The Phyre2 web portal for protein modeling, prediction and analysis. Nature Protocols 10(6), 845-858.
| Crossref | Google Scholar | PubMed |

Kumar S, Stecher G, Li M, Knyaz C, Tamura K (2018) MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology and Evolution 35(6), 1547-1549.
| Crossref | Google Scholar | PubMed |

Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. Journal of Molecular Biology 157(1), 105-132.
| Crossref | Google Scholar | PubMed |

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4), 402-408.
| Crossref | Google Scholar | PubMed |

Meena PD, Awasthi RP, Chattopadhyay C, Kolte SJ, Kumar A (2016) Alternaria blight: a chronic disease in rapeseed-mustard. Journal of Oilseed Brassica 2010(1), 1-11.
| Google Scholar |

Mitra PP, Loque D (2014) Histochemical staining of Arabidopsis thaliana secondary cell wall elements. Journal of Visualized Experiments [87] e51381.
| Crossref | Google Scholar |

Moura JCMS, Bonine CAV, Viana JdOF, Dornelas MC, Mazzafera P (2010) Abiotic and biotic stresses and changes in the lignin content and composition in plants. Journal of Integrative Plant Biology 52(4), 360-376.
| Crossref | Google Scholar | PubMed |

Ni Z, Lü M, Fan L (2010) Cloning and characterization of CCoAOMT gene from Gossypium hirsutum L. Acta Botanica Boreali-Occidentalia Sinica 30(6), 1083-1091.
| Google Scholar |

Pawelec A, Dubourg C, Briard M (2006) Evaluation of carrot resistance to Alternaria leaf blight in controlled environments. Plant Pathology 55(1), 68-72.
| Crossref | Google Scholar |

Pearson K (1896) Mathematical contributions to the theory of evolution. II. On the theory of contiguity. Philosophical Transactions of the Royal Society of London. Series A, Containing Papers of a Mathematical or Physical Character 187, 253-318.
| Google Scholar |

Rakoczy M, Femiak I, Alejska M, Figlerowicz M, Podkowinski J (2018) Sorghum CCoAOMT and CCoAOMT-like gene evolution, structure, expression and the role of conserved amino acids in protein activity. Molecular Genetics and Genomics 293(5), 1077-1089.
| Crossref | Google Scholar | PubMed |

Riaz MW, Yousaf MI, Hussain Q, Yasir M, Sajjad M, Shah L (2023) Role of lignin in wheat plant for the enhancement of resistance against lodging and biotic and abiotic stresses. Stresses 3(2), 434-453.
| Crossref | Google Scholar |

Sharma A, Sambasivam V, Shukla P, Rampuria S, Kirti PB (2020) An in vitro generated variant of Tephrosia villosa defensin (α-TvD1) enhances biotic stress tolerance in transgenic tobacco. Journal of Plant Pathology 102, 1133-1143.
| Crossref | Google Scholar |

Simms D, Cizdziel PE, Chomczynski P (1993) TRIzol: a new reagent for optimal single-step isolation of RNA. Focus 15(4), 532-535.
| Google Scholar |

Song J-L, Wang Z-Y, Wang Y-H, Du J, Wang C-Y, Zhang X-Q, Chen S, Huang X-L, Xie X-M, Zhong T-X (2022) Overexpression of Pennisetum purpureum CCoAOMT contributes to lignin deposition and drought tolerance by promoting the accumulation of flavonoids in transgenic tobacco. Frontiers in Plant Science 13, 884456.
| Crossref | Google Scholar | PubMed |

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25(24), 4876-4882.
| Crossref | Google Scholar | PubMed |

Walker AM, Sattler SA, Regner M, Jones JP, Ralph J, Vermerris W, Sattler SE, Kang C (2016) The structure and catalytic mechanism of Sorghum bicolor caffeoyl-CoA O-methyltransferase. Plant Physiology 172(1), 78-92.
| Crossref | Google Scholar | PubMed |

Wang M, Zhu X, Wang K, Lu C, Luo M, Shan T, Zhang Z (2018) A wheat caffeic acid 3-O-methyltransferase TaCOMT-3D positively contributes to both resistance to sharp eyespot disease and stem mechanical strength. Scientific Reports 8(1), 6543.
| Crossref | Google Scholar |

Wei L, Zhao X, Gu X, Peng J, Song W, Deng B, Cao Y, Hu S (2023) Genome-wide identification and expression analysis of Dendrocalamus farinosus CCoAOMT gene family and the role of DfCCoAOMT14 involved in lignin synthesis. International Journal of Molecular Sciences 24(10), 8965.
| Crossref | Google Scholar | PubMed |

Xiong XP, Sun SC, Zhu QH, Zhang XY, Li YJ, Liu F, Xue F, Sun J (2021) The cotton lignin biosynthetic gene Gh4CL30 regulates lignification and phenolic content and contributes to Verticillium wilt resistance. Molecular Plant-Microbe Interactions 34(3), 240-254.
| Crossref | Google Scholar | PubMed |

Yang Q, He Y, Kabahuma M, Chaya T, Kelly A, Borrego E, Bian Y, Kasmi FE, Yang L, Teixeira P, Kolkman J, Nelson R, Kolomiets M, Dangl JL, Wisser R, Caplan J, Li X, Lauter N, Balint-Kurti P (2017) A gene encoding maize caffeoyl-CoA O-methyltransferase confers quantitative resistance to multiple pathogens. Nature Genetics 49(9), 1364-1372.
| Crossref | Google Scholar | PubMed |

Yang G, Pan W, Zhang R, Pan Y, Guo Q, Song W, Nie X (2021) Genome-wide identification and characterization of caffeoyl-coenzyme A O-methyltransferase genes related to the Fusarium head blight response in wheat. BMC Genomics 22(1), 504.
| Crossref | Google Scholar |

Zhong R, Morrison III W, Himmelsbach DS, Poole FL, Ye ZH (2000) Essential role of caffeoyl coenzyme A-O-methyltransferase in lignin biosynthesis in woody poplar plants. Plant Physiology 124(2), 563-578.
| Crossref | Google Scholar | PubMed |