Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
Shopping Cart: ( empty )
You are here: Home > Journals > FP > FP25049
RESEARCH ARTICLE
Contents Vol 52(7)

Discovery of protein interactors of SUGARWINs in Saccharum officinarum using cDNA library screening

Aqsa Parvaiz https://orcid.org/0000-0001-8180-2168 A # , Muhammad Mubashar Zafar https://orcid.org/0000-0002-5664-138X B C # , Faiz Ahmad Joyia D , Sultana Anwar E , Andrei Smertenko F , Hira Kamal https://orcid.org/0000-0001-6503-2167 G , Saba Zafar https://orcid.org/0000-0001-7317-0084 A , Mishal Subhan https://orcid.org/0000-0002-6797-9175 H , Muhammad Sarwar Khan D , Mahmoud F. Seleiman I , Majed A. Alotaibi https://orcid.org/0009-0000-3402-2476 I , Sezai Ercisli J , Ghulam Mustafa D * and Xuefei Jiang B *
+ Author Affiliations
- Author Affiliations

A Department of Biochemistry and Biotechnology, The Women University Multan, Multan, Pakistan. Email: aqsapervaiz333@gmail.com; saba.6498@wum.edu.pk

B Sanya Institute of Breeding and Multiplication/School of Tropical Agriculture and Forestry, Hainan University, Sanya, China.

C Department of Plant Breeding and Genetics, University of Agriculture Faisalabad, Faisalabad, Pakistan. Email: m.mubasharzafar@gmail.com

D Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture Faisalabad, Faisalabad, Pakistan. Email: faizahmad1980@gmail.com

E Department of Plant and Microbial Biology, University of California Berkeley, Berkeley, CA, USA. Email: sultana.anwar@yahoo.com

F Institute of Biological Chemistry (IBC), Washington State University, Pullman, WA, USA. Email: andrei.smertenko@wsu.edu

G Department of Plant Pathology, Washington State University, Pullman, WA, USA. Email: hira.kamal111@gmail.com

H Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan. Email: mishalsubhan@gmail.com

I Department of Plant Production, College of Food and Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia. Email: mseleiman@ksu.edu.sa

J Department of Horticulture, Faculty of Agriculture, Ataturk University, Erzurum, Turkey. Email: sercisli@atauni.edu.tr

* Correspondence to: drmustafa8@gmail.com, xuefei.jiang@hainanu.edu.cn

# These authors contributed equally to this work

Handling Editor: Sajid Fiaz

Functional Plant Biology 52, FP25049 https://doi.org/10.1071/FP25049
Submitted: 21 February 2025  Accepted: 23 June 2025  Published: 14 July 2025

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

Abstract

Sugarcane holds considerable commercial significance due to its role as the primary source of sugar and its potential as a global biofuel resource. Fungal pathogens and insect pests present significant challenges to the cultivation of this crop, leading to substantial reductions in crop yield and sugar recovery. In response to pathogen infection, plants initiate their defense mechanisms, which involve the upregulation of pathogenesis-related proteins such as chitinase, glucanase, and chitosanase. SUGARWINs refer to a group of PR-4 proteins that are associated with the defense mechanisms of sugarcane against phytopathogens. Their gene expression is induced in response to wounds caused by Diatraea saccharalis larvae and diseases caused by fungal pathogens such Colletothricum falcatum and Fusarium verticillioides. We report the finding of some other proteins that interact with SUGARWINs and may also have a role in the defense against fungal diseases. The sugarcane cDNA library was screened against SUGARWIN1 and SUGARWIN2 proteins to find possible interactors. A strong interaction of both SUGARWIN1 and SUGARWIN2 was observed with oxygen evolving enhancer protein 1 and synaptotagmin 1. These interactions were further validated by BiFC (biomolecular fluorescence complementation) assay. For further molecular characterization, subcellular localization studies of SUGARWINs and interactor proteins were conducted by translational fusion with green fluorescent protein.

Keywords: biomolecular fluorescence complementation, cDNA library screening, Colletotrichum falcatum, pathogenesis-related proteins, plant defence mechanisms, plant immunity signaling, Saccharum officinarum, subcellular localisation, SUGARWINs.

References

Afghan S, Khan ME, Verma KK, Nikpay A (2024) Economic importance and yield potential of sugarcane in Pakistan. In ‘Sugarcane cultivation and management’. (Eds KK Verma, X-P Song, M Singh, R de Mello Prado, J-M Wu, Y-R Li) pp. 263–291. (Apple Academic Press)

Agnihotri V, Madan VJSC (1989) Changes in carbohydrates and nucleic acids in sugar cane varieties infected with both leaf scald and red rot. CABI, Record No. 19901140286.

Ahmad M, Ali R, Fasihi S (1986) Effect of different infection levels of red rot of sugarcane on cane weight and juice quality. Journal of Agricultural Research 24(2), 129-131.
| Google Scholar |

Ahmad M, Sadiq S, Asghar S, Hayat F, Hussain N, Hameed M, Karamat U, Shaheen S, Khan MS, Joyia FA (2025) Advancing in vitro propagation of true-to-type lines of Amaranthus spp. employing SSR-based genetic evaluation. Agrobiological Records 19, 78-91.
| Google Scholar |

Ahmed SR, Anwar Z, Shahbaz U, Skalicky M, Ijaz A, Tariq MS, Zulfiqar U, Brestic M, Alabdallah NM, Alsubeie MS, Mujtaba H, Saeed AM, Zahra T, Hasan MM, Firdous H, Razzaq A, Zafar MM (2023) Potential role of silicon in plants against biotic and abiotic stresses. Silicon 15(7), 3283-3303.
| Crossref | Google Scholar |

Ali M, Shaukat F, Khan W, Syed A, Maqsood J, Kamal H, Ercisli S, Balijagic J, Ullah R, Bari A, Anwar Z, Ijaz A, Kashif M, Khalid MN, Mustafa HSB, Zafar MM (2023) Microsatellite-based diversity analysis and the development of core-set germplasm in Pakistani barley lines. Cellular and Molecular Biology 69(10), 100-108.
| Crossref | Google Scholar | PubMed |

Alotibi YS (2024) A systematic review on adoption of biodegradable mulches among farmers. International Journal of Agriculture and Biosciences 13(4), 777-783.
| Google Scholar |

Aristya GR, Nabillah SAA, Kasiamdari RS, Damaiyani J, Prabowo H (2024) Phylogenetic and genetic variation of sugarcane (Saccharum spp.) from Island of Java, Indonesia based on trnK chloroplast gene. International Journal of Agriculture and Biosciences 13(2), 157-166.
| Google Scholar |

Berndsen CE, Storm AR, Sardelli AM, Hossain SR, Clermont KR, McFather LM, Connor MA, Monroe JD (2025) The pseudoenzyme β-Amylase9 from arabidopsis activates α-Amylase3: a possible mechanism to promote stress-induced starch degradation. Proteins 93(6), 1189-1201.
| Crossref | Google Scholar |

De Silva G, Silva M, Carvalho P (1977) Preliminary studies on invertases of sugarcane stalks inoculated with C. falcatum Went. In ‘Proceedings of the International Society of Sugar Cane Technologists’, 9–25 January 1977, Sao Paulo, Brazil. (CABI)

Dimobe K, Zoungrana BJ-B, Yoni M, Thiombiano A (2025) Agroforestry’s contribution to sustainable soil fertility, livelihoods, and carbon sequestration in Sub-Saharan Africa: a systematic review. International Journal of Agriculture and Biosciences 14(3), 436-446.
| Crossref | Google Scholar |

Du M, Zhou X, Yang X (2025) Identification of interaction between geminivirus and plant proteins by bimolecular fluorescence complementation (BiFC). In ‘Geminiviruses: methods and protocols’. (Eds FM Zerbini, E Fiallo-Olivé, J Navas-Castillo) pp. 227–234. (Humana: New York, NY, USA)

Fañanás-Pueyo I, Anhel AM, Goñi-Moreno Á, Oñate-Sánchez L, Carrera-Castaño G (2024) Workflow to select functional promoter DNA baits and screen arrayed gene libraries in yeast. Current protocols 4(11), e70059.
| Crossref | Google Scholar | PubMed |

Fields S, Song O-k (1989) A novel genetic system to detect protein–protein interactions. Nature 340(6230), 245-246.
| Crossref | Google Scholar | PubMed |

Franco FP (2017) Unraveling sugarcane-Diatraea saccharalis-opportunistic fungi interaction in sugarcane. Doctor thesis, Universidade de São Paulo.

Franco FP, Santiago AC, Henrique-Silva F, de Castro PA, Goldman GH, Moura DS, Silva-Filho MC (2014) The sugarcane defense protein SUGARWIN2 causes cell death in Colletotrichum falcatum but not in non-pathogenic fungi. PLoS ONE 9(3), e91159.
| Crossref | Google Scholar | PubMed |

Galletta BJ, Rusan NM (2015) A yeast two-hybrid approach for probing protein–protein interactions at the centrosome. In ‘Centrosome & Centriole’. Methods in Cell Biology. (Eds R Basto, K Oegema) vol. 129, pp. 251–277. (Elsevier)

Hein MY, Peng D, Todorova V, McCarthy F, Kim K, Liu C, Savy L, Januel C, Baltazar-Nunez R, Sekhar M, Vaid S, Bax S, Vangipuram M, Burgess J, Njoya L, Wang E, Ivanov IE, Byrum JR, Pradeep S, Gonzalez CG, Aniseia Y, Creery JS, McMorrow AH, Sunshine S, Yeung-Levy S, DeFelice BC, Mehta SB, Itzhak DN, Elias JE, Leonetti MD (2025) Global organelle profiling reveals subcellular localization and remodeling at proteome scale. Cell 188(4), 1137-1155.e20.
| Crossref | Google Scholar | PubMed |

Holkar SK, Balasubramaniam P, Kumar A, Kadirvel N, Shingote PR, Chhabra ML, Kumar S, Kumar P, Viswanathan R, Jain RK, Pathak AD (2020) Present status and future management strategies for Sugarcane yellow leaf virus: a major constraint to the global sugarcane production. The Plant Pathology Journal 36(6), 536-557.
| Crossref | Google Scholar | PubMed |

Horstman A, Nougalli Tonaco IA, Boutilier K, Immink RGH (2014) A cautionary note on the use of split-YFP/BiFC in plant protein–protein interaction studies. International Journal of Molecular Sciences 15(6), 9628-9643.
| Crossref | Google Scholar | PubMed |

Hwang H-H, Gelvin SB (2004) Plant proteins that interact with VirB2, the Agrobacterium tumefaciens pilin protein, mediate plant transformation. The Plant Cell 16(11), 3148-3167.
| Crossref | Google Scholar | PubMed |

Kamal H, Zafar MM, Parvaiz A, Razzaq A, Elhindi KM, Ercisli S, Qiao F, Jiang X (2024) Gossypium hirsutum calmodulin-like protein (CML 11) interaction with geminivirus encoded protein using bioinformatics and molecular techniques. International Journal of Biological Macromolecules 269, 132095.
| Crossref | Google Scholar | PubMed |

Karban R, Baldwin IT (2007) ‘Induced responses to herbivory.’ (University of Chicago Press)

Medeiros AH, Franco FP, Matos JL, de Castro PA, Santos-Silva LK, Henrique-Silva F, Goldman GH, Moura DS, Silva-Filho MC (2012) Sugarwin: a sugarcane insect-induced gene with antipathogenic activity. Molecular Plant-Microbe Interactions 25(5), 613-624.
| Crossref | Google Scholar | PubMed |

Munir H, Abbas S, Sohail MT (2023) Nurturing the crop: recent innovations in vegetable diseases management. Agrobiological Records 13, 101-107.
| Crossref | Google Scholar |

Parvaiz A, Mustafa G, Khan HMWA, Joyia FA, Niazi AK, Anwar S, Khan MS (2019) Field evaluation ratified by transcript and computational analyses unveils myco-protective role of SUGARWIN proteins in sugarcane. 3 Biotech 9, 377.
| Crossref | Google Scholar |

Parvaiz A, Mustafa G, Khan MS, Ali MA (2021) Over-expression of endogenous SUGARWIN genes exalted tolerance against Colletotrichum infection in sugarcane. Plants 10(5), 869.
| Crossref | Google Scholar | PubMed |

Parvaiz A, Joyia FA, Saeed M, Azwar M, Khan MS, Mustafa G (2023) Defense-related proteins in sugarcane and their role in disease resistance: molecular advancements and beyond. In ‘Agro-industrial perspectives on sugarcane production under environmental stress’. (Eds KK Verma, XP Song, VD Rajput, S Solomon, YR Li, GP Rao) pp. 171–192. (Springer: Singapore)

Peng C, Chang L, Yang Q, Tong Z, Wang D, Tan Y, Sun Y, Yi X, Ding G, Xiao J, Zhang Y, Wang X (2019) Comparative physiological and proteomic analyses of the chloroplasts in halophyte Sesuvium portulacastrum under differential salt conditions. Journal of Plant Physiology 232, 141-150.
| Crossref | Google Scholar | PubMed |

Printen JA, Sprague GF, Jr. (1994) Protein-protein interactions in the yeast pheromone response pathway: Ste5p interacts with all members of the MAP kinase cascade. Genetics 138(3), 609-619.
| Crossref | Google Scholar | PubMed |

Prompreing K, Saikamwong N, Aekram S (2025) Enhancing agricultural productivity and innovation: deep learning for citrus disease classification in Thai orchards. International Journal of Agriculture and Biosciences 14(2), 283-288.
| Google Scholar |

Rauf S, Shahid A, Faizan M, Khalid MN, Amjad I (2024) CRISPER/CAS: a potential tool for genomes editing. Agrobiological Records 15, 13-23.
| Crossref | Google Scholar |

Reymond P, Bodenhausen N, Van Poecke RMP, Krishnamurthy V, Dicke M, Farmer EE (2004) A conserved transcript pattern in response to a specialist and a generalist herbivore. The Plant Cell 16(11), 3132-3147.
| Crossref | Google Scholar | PubMed |

Ricaud C, Egan B, Gillaspie A, Hughes C (2012) ‘Diseases of sugarcane: major diseases.’ (Elsevier)

Rocha FR, Papini-Terzi FS, Nishiyama MY, Vencio RZ, Vicentini R, Duarte RD, de Rosa VE, Vinagre F, Barsalobres C, Medeiros AH, Rodrigues FA, Ulian EC, Zingaretti SM, Galbiatti JA, Almeida RS, Figueira AV, Hemerly AS, Silva-Filho MC, Menossi M, Souza GM (2007) Signal transduction-related responses to phytohormones and environmental challenges in sugarcane. BMC Genomics 8, 71.
| Crossref | Google Scholar |

Rodrigues FA, de Laia ML, Zingaretti SM (2009) Analysis of gene expression profiles under water stress in tolerant and sensitive sugarcane plants. Plant Science 176(2), 286-302.
| Crossref | Google Scholar |

Rott P (2000) ‘A guide to sugarcane diseases.’ (Editions Quae)

Saeed MZ, Hayat H, Shafiq F, Tareen W-ul-H (2023) Assessing the prospects and challenges of organic agriculture in the pothwar Region of Punjab, Pakistan. International Journal of Agriculture and Biosciences 12(1), 1-7.
| Google Scholar |

Shabbir MA, Iftikhar Y, Mubeen M, Zeshan MA, Jun Y, Almutairi LA, Ahmed AE (2025) Effect of Okra Yellow Vein Mosaic Disease on biochemical components of okra plants. Pakistan Journal of Agricultural Sciences 62, 291-299.
| Crossref | Google Scholar |

Shahid M, Shaukat F, Shahid A, Sohail A, Nadeem M (2023) Biopesticides: a potential solution for the management of insect pests. Agrobiological Records 13, 7-15.
| Crossref | Google Scholar |

Souza TP, Dias RO, Silva-Filho MC (2017) Defense-related proteins involved in sugarcane responses to biotic stress. Genetics and Molecular Biology 40, 360-372.
| Crossref | Google Scholar | PubMed |

Stynen B, Tournu H, Tavernier J, Van Dijck P (2012) Diversity in genetic in vivo methods for protein–protein interaction studies: from the yeast two-hybrid system to the mammalian split-luciferase system. Microbiology and Molecular Biology Reviews 76(2), 331-382.
| Crossref | Google Scholar | PubMed |

Suo J, Zhang H, Zhao Q, Zhang N, Zhang Y, Li Y, Song B, Yu J, Cao J, Wang T, Luo J, Guo L, Ma J, Zhang X, She Y, Peng L, Ma W, Guo S, Miao Y, Chen S, Qin Z, Dai S (2020) Na2CO3-responsive photosynthetic and ROS scavenging mechanisms in chloroplasts of alkaligrass revealed by phosphoproteomics. Genomics, Proteomics and Bioinformatics 18(3), 271-288.
| Crossref | Google Scholar | PubMed |

Viswanathan R (2012) Molecular basis of red rot resistance in sugarcane. Functional Plant Science and Biotechnology 6(2), 40-50.
| Google Scholar |

Viswanathan R (2021) Red rot of sugarcane (Colletotrichum falcatum Went). CABI Reviews. 10.1079/PAVSNNR202116023

Viswanathan R, Samiyappan R (1999) Red rot disease in sugarcane: a major constraint for the Indian sugar industry. Sugar Cane 5, 9-15.
| Google Scholar |

Walling LL (2000) The myriad plant responses to herbivores. Journal of Plant Growth Regulation 19, 195-216.
| Crossref | Google Scholar | PubMed |

Wu J, Baldwin IT (2009) Herbivory-induced signalling in plants: perception and action. Plant, Cell & Environment 32(9), 1161-1174.
| Crossref | Google Scholar | PubMed |

Wu L, Han Z, Wang S, Wang X, Sun A, Zu X, Chen Y (2013) Comparative proteomic analysis of the plant–virus interaction in resistant and susceptible ecotypes of maize infected with sugarcane mosaic virus. Journal of Proteomics 89, 124-140.
| Crossref | Google Scholar | PubMed |