Ricin detoxification in castor (Ricinus communis L.) seeds: Current advances, integrated strategies, and future directions
RICIN DETOXIFICATION IN CASTOR : ADVANCES, INTEGRATED STRATEGIES, AND FUTURE DIRECTIONS
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Keywords:
Castor bean, Cas9, Detoxification, Genome editing, RCA. Ricin, RNAiAbstract
Castor bean (Ricinus communis L.) is a key industrial crop valued for its non-edible oil rich in ricinoleic acid. However, the presence of ricin - a type II ribosome-inactivating protein (RIP)-in the seed endosperm remains a major bottleneck for valorizing the protein-rich de-oiled cake (~20-25% protein) as animal feed. Ricin is highly toxic, with an LD50 of ~1-5 µg/kg in humans, and poses significant risks to human and animal health, thereby limiting the crop's commercial potential. Traditional detoxification methods, including physical (thermal), chemical (alkali/acid), and enzymatic treatments offer limited efficacy, often compromising protein quality or leaving toxic residues. Recent advances in plant genomics and molecular biology have facilitated the identification and characterization of ricin and RCA gene families, enabling targeted gene silencing and editing approaches. RNA interference (RNAi), antisense suppression, and CRISPR/Cas9-mediated knockouts have demonstrated promising results in reducing ricin expression while maintaining normal seed development and oil composition. Novel strategies such as promoter engineering, tissue-specific expression control, and base editing are emerging as powerful tools for generating transgene-free, low-ricin castor lines. Additionally, microbial degradation using enzymes and biotransformation pathways offers eco-friendly and scalable alternatives. This review discusses the current knowledge on the structure and biosynthesis of ricin, its mechanisms of toxicity, and highlights the latest strategies and technological interventions aimed at detoxifying castor bean seeds. We also evaluate the advantages, limitations, and regulatory challenges of these approaches and propose future research directions for sustainable, safe, and economically viable detoxification of castor products.
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Akande, T O, Odunsi A A and Akinfala E O 2016. A review of nutritional and toxicological implications of castor bean (Ricinus communis L.) meal in animal feeding systems. Journal of Animal Physiology and Animal Nutrition, 100(2): 201-210.
Anjani K 2014. A re-evaluation of castor (Ricinus communis L.) as a crop plant. CABI Reviews, 1-21.
Anandan S, Anil Kumar G K, Ghosh J and Ramachandra K S 2005. Effect of different physical and chemical treatments on detoxification of ricin in castor cake. Animal Feed Science and Technology, 120: 159-168.
Ashfaq M A, Reddy P S, Anil-Kumar C, Selvaraj V M and Kumar V D 2018. Ricin and RCA- The enemies within castor (Ricinus communis L.): A perspective on their biogenesis, mechanism of action, detection methods and detoxification strategies. In: C Kole and P Rabinowicz (Eds.), The castor bean genome. Springer Nature, Switzerland, pp. 215-236.
Ashfaq M A, Reddy P S, Kumar CA, Kirti P B and Kumar VD 2010. Towards functional analysis of ricin promoters. Journal of Oilseeds Research, 27: 12-14.
Ashfaq M A, Rao N N, Kirti P B and Kumar V D 2009. Isolation of ricin promoters from castor (Ricinus communis L.). Journal of Oilseeds Research, 26: 208-210.
Bally J, Fishilevich E, Bowling AJ, Pence HE, Narva KE and Waterhouse PM 2018. Improved insect-proofing: expressing double-stranded RNA in chloroplasts. Pest Management Science, 74(8): 1751-1758.
Bass H W, Krawetz J E, Obrian G R, Zinselmeier C, Jeffrey E, Habben J E and Boston R S 2004. Maize ribosome-inactivating proteins (RIPs) with distinct expression patterns have similar requirements for proenzyme activation, Journal of Experimental Botany, 55(406): 2219-2233. https://doi.org/10.1093/jxb/erh243.
Borphukan B, Khatun M, Fartyal D, James D and Reddy MK 2025. A Gemini Virus-Derived Autonomously Replicating System for HDR-Mediated Genome Editing of the EPSP Synthase Gene in Indica Rice. Plants-Basel, 14(3): 477. doi: 10.3390/plants14030477.
Brandon D L, McKeon T A, Patfield S A and He X 2016. Analysis of castor by ELISAs that distinguish Ricin and Ricinus communis agglutinin (RCA). Journal of theAmerican Oil Chemists' Society, 93: 359-363.
Brown AP, Kroon JT, Swarbreck D, Febrer M, Larson TR, Graham IA, Caccamo M and Slabas AR 2012. Tissue-specific whole transcriptome sequencing in castor, directed at understanding triacylglycerol lipid biosynthetic pathways. PLoS One, 7(2): e30100.
Chan AP, Crabtree J, Zhao Q, Lorenzi H, Orvis J, Puiu D, Melake-Berhan A, Jones KM, Redman J, Chen G, Cahoon EB, Gedil M, Stanke M, Haas BJ, Wortman JR, Fraser-Liggett CM, Ravel J and Rabinowicz PD 2010. Draft genome sequence of the oilseed species Ricinus communis. Nature Biotechnology, 28(9): 951-6.
ChaudhryB, Müller-Uri F, Cameron-Mills V, Gough S, Simpson D, Skriver K and Mundy J 1994. The barley 60 kDa jasmonate-induced protein (JIP60) is a novel ribosome-inactivating protein. The Plant Journal, 6(6): 815-24.
Chen S, Gistelinck K, Verbeke I and Van Damme EJM 2022. Differential effects of the recombinant type 1 ribosome-inactivating protein, OsRIP1, on growth of PSB-D and BY-2 cells. Front Plant Science, 29(13): 1019591. doi: 10.3389/fpls.2022.1019591.
Darby, Shauna M, Mark L Miller and Ralph O Allen 2001. Forensic determination of ricin and the alkaloid marker ricinine from castor bean extracts. Journal of Forensic Sciences, 46(5): 1033-1042.
De Sousa NL, Cabral GB and Aragão FJL 2022. Herbicide tolerance and gene silencing stability over generations in the ricin bio-detoxicated castor bean. Journal of Genetic Engineering and Biotechnology, 20(1): 16. doi: 10.1186/s43141-022-00303-w.
Dua P, Yoo J W, Kim S and Lee D K 2011. Modified siRNA structure with a single nucleotide bulge overcomes conventional siRNA-mediated off-target silencing. Molecular Therapy, 19(9): 1676-1687.
Endo Y and Tsurugi, K 1988. The RNA N-Glycosidase Activity of Ricicn A-chain. The Characteristics of the Enzymatic Activity of Ricin A-chain with Ribosomes and with rRNA. The Journal of Biological Chemistry, 263: 8735-9. 10.1016/S0021-9258(18)68367-X.
Fernandes KV, Deus-de-Oliveira N, Godoy MG, Guimarães ZA, Nascimento VV, Melo EJ, Freire DM, Dansa-Petretski M and Machado OL 2012. Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds. Brazilian Journal of Medical and Biological Research, 45(11):1002-10. doi: 10.1590/s0100-879x2012007500132.
Frankel A and Roberts RW 2003. In vitro selection for sense codon suppression. RNA Journal, 9(7): 780-6. doi: 10.1261/rna.5350303.
Gardner H K Jr., D'Aquin EL, Koltun SP, McCourtney EJ, Vix HLE and Gastrock EA 1960. Detoxification and deallergenization of castor beans. Journal of the American Oil Chemistry Society, 37: 142-148. https://doi.org/10.1007/BF02644933.
Ghosh A, Shekhawat UKS, Ganapathi TR and Bapat VA 2012. Analysis of banana fruit-specific promoters using transient expression in embryogenic cells of banana cultivar Robusta (AAA Group). Journal of Plant Biochemistry and Biotechnology, 21(2): 189-197.
Hartley MR and Lord JM 2004. Cytotoxic ribosome-inactivating lectins from plants. Biochimica Biophysica Acta, 1701(1-2): 1-14. doi: 10.1016/j.bbapap.2004.06.004. PMID: 15450171.
Iglesias R, Russo R, Landi N, Valletta M, Chambery A, Di Maro A, Bolognesi A, Ferreras JM, Citores L 2022. Structure and Biological Properties of Ribosome-Inactivating Proteins and Lectins from Elder (Sambucus nigra L.) Leaves. Toxins-Basel, 14(9): 611.
Komor A, Kim Y and Packer M 2016. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533: 420-424. https://doi.org/10.1038/nature17946.
Kumaraswamy H H 2022. Biotechnological Approaches for Genetic Improvement of Castor Bean (Ricinus communis L.). In: Gosal, S.S., Wani, S.H. (eds) Accelerated Plant Breeding, Springer, 4.
Lakshmidevi G, Bhavani K, Sreevathsa Rohini and Kumar V 2023. Agrobacterium-mediated in planta transformation in castor (Ricinus communis L.). Journal of Oilseeds Research, 35(3). Doi: 10.56739/jor.v35i3.137454.
Lee-Huang, Sylvia, Philip L. Huang, Hsiang-Fu Kung, Bao-Qun Li, Paul L. Huang, Peter Huang, Henry I. Huang, and Hao-Chia Chen 1991. TAP 29: An anti-human immunodeficiency virus protein from Trichosanthes Kirilowii that is nontoxic to intact cells. Proceedings of the National Academy of Sciences of the United States of America, 88(15): 6570-6574.
Lima M V S, Bagaldo A R, Muller M, Pinheiro E G, Almeida B J, Mugab L C, Araujo F L, Bezerra L R, Oliveira R L 2020. Intake, digestibility, ingestive behavior, production, and composition of goat milk supplemented with detoxified castor bean meal added urea as a replacement of soybean meal. Tropical Animal Health Production, 52(4): 2135-2143.
Lord PW, Stevens RD, Brass A, Goble CA 2003. Semantic similarity measures as tools for exploring the gene ontology. Pacific Symposium Biocomputing, 601-612.
Loss-Morais G, Turchetto-Zolet AC, Etges M, Cagliari A, Körbes AP, Maraschin Fdos S, Margis-Pinheiro M, Margis R 2013. Analysis of castor bean ribosome-inactivating proteins and their gene expression during seed development. Genetics Molecular Biology, 36(1): 74-86.
Lu J, Pan C, Fan W, Liu W, Zhao H, Li D, Wang S, Hu L, He B, Qian K, Qin R, Ruan J, Lin Q, Lü S, Cui P 2022. A Chromosome-level Genome Assembly of Wild Castor Provides New Insights into its Adaptive Evolution in the Tropical Desert. Genomics Proteomics Bioinformatics, 20(1): 42-59.
Melo, Walber, Santos, Alexandre, Maria Lídia and Pereira Jr Nei 2008. Acid and Enzymatic Hydrolysis of the Residue from Castor Bean (Ricinus communis L.) Oil Extraction for Ethanol Production: Detoxification and Biodiesel Process Integration. Journal of The Brazilian Chemical Society - JBCS, 19.
Miller J C, Patil D P, Xia D F, Paine C B, Fauser F, Richards H W, Shivak D A, Bendaña YR, Hinkley S J, Scarlott N A and Lam S C 2019. Enhancing gene editing specificity by attenuating DNA cleavage kinetics. Nature Biotechnology, 37(8): 945-952.
Ogunniyi and David S 2006. Castor oil: a vital industrial raw material. Bioresource Technology, 97(9): 1086-1091.
Oliveira André, Oliveira M R C, Campos José, Lana Rogério, Machado Olga, Retamal CA, Detmann Edenio and Valadares Filho Sebastiao 2000. In vitro ruminal degradation of ricin and its effect on microbial growth. Animal Feed Science and Technology, 10.1016/j.anifeedsci.2010.01.006.
Ready MP, Kim Y and Robertus JD 1991. Site-directed mutagenesis of ricin A-chain and implications for the mechanism of action. Proteins, 10:270-8.
ReddyPS, Ashfaq MA, Kumar CA, Kirti PB and Kumar VD 2009 Development of a transgenic tobacco model system as a prelude
to identify the efficient PTGS technology for silencing ricin and RCA in castor, Ricinus communis L. Journal of Oilseeds Research, 26: 199-202.
Reddy P S, Sharan S T and Kumar V D 2010. Transgenic tobacco - as a model for identifying the most efficient PTGS technology at silencing ricin and RCA in castor (Ricinus communis L). Journal of Oilseeds Research, 27: 7-11.
Rivarola M, Foster JT, Chan AP, Williams AL, Rice DW, Liu X, Melake-Berhan A, Huot Creasy H, Puiu D, Rosovitz MJ, Khouri HM, Beckstrom-Sternberg SM, Allan GJ, Keim P, Ravel J and Rabinowicz PD 2011. Castor bean organelle genome sequencing and worldwide genetic diversity analysis. PLoS One, 6(7): e21743. doi: 10.1371/journal.pone.0021743.
Saikumar K, SwamyHHK and Kumar VD 2009 Enemy of enemy: In silico designed amiRNAs to silence ricin genes in castor, Ricinus communis L. Journal of Oilseeds Research, 26: 188-190.
Salim R, Nehvi IB, Mir RA, Tyagi A, Ali S, Bhat OM., 2023 A review on anti-nutritional factors: unraveling the natural gateways to human health. Frontiers in Nutrition, 10: 1215873.
Severino Liv, Auld Dick, Baldanzi Marco, Cândido M, Chen Grace, Crosby William, Tan D, He Xiaohua, Parvathaneni Lakshmamma, Lavanya Ch, Machado Olga, Mielke Thomas, Milani Máira, Miller Travis, Morris J, Morse Stephen, Navas Alejandro, Soares Dartanha, Sofiatti Valdinei and Zieler Helge 2012. A Review on the Challenges for Increased Production of Castor. Agronomy Journal, 104: 853.
Sousa NL, Cabral GB, Vieira PM, Baldoni AB and Aragão FJL 2017. Bio-detoxification of ricin in castor bean (Ricinus communis L.) seeds. Science Reporter, 7(1): 15385.
Stirpe F and Battelli MG 2006. Ribosome-inactivating proteins: progress and problems. Cell Molecular Life Science, 63(16): 1850-1866.
Tregear, James W and Lynne M Roberts 1992. The lectin gene family of Ricinus communis: cloning of a functional ricin gene and three lectin pseudogenes. Plant Molecular Biology, 18: 515-525.
Tumer and Nilgun E 2019. Introduction to the toxins special issue "ricin toxins". Toxins, 12(1): 13.
Usha Kiran B, Kumar V D, Kumaraswamy HH and Sujatha M 2020. In vitro regeneration of castor (Ricinus communis L.): In vitro regeneration of castor (Ricinus communis L.). Journal of Oilseeds Research, 37(Special issue).
Walsh MJ, Dodd JE and Hautbergue GM 2013. Ribosome inactivating proteins: potent poisons and molecular tools. Virulence, 4(8):774-784.
Xie Y, Zhang W, Li Y, Wang M, Cerny R L, Shen Y and Du L 2011. Transformation of Fusarium verticillioides with a polyketide gene cluster isolated from a fungal endophyte activates the biosynthesis of fusaric acid. Mycology, 2(1): 24-29.
Zhang F, Song G and Tian Y 2019. Anti-CRISPRs: The natural inhibitors for CRISPR-Cas systems. Animal Model and Experimental Medicine, 2(2): 69-75.
Zhu F, Zhou YK, Ji ZL and Chen XR 2018. The Plant Ribosome-Inactivating Proteins Play Important Roles in Defense against Pathogens and Insect Pest Attacks. Front Plant Science, 9:146.doi: 10.3389/fpls.