Abstract
Some edible Leguminous are toxic when raw, and the Chinese are particularly fond of beans, so Leguminous poisoning is very common in China. Rapid and accurate identification of poisoned species and determination of their toxic components would better assist physicians in treating patients. However, traditional morphology-based identification methods possess many limitations. DNA barcoding technique is a new species identification technique developed in recent years, which is expected to make up for the shortcomings of traditional morphological identification. In this study, a comprehensive evaluation system based on DNA barcoding and ELISA kits was attempted. A total of 30 Leguminous toxic plants were collected, involving 9 genera and 10 species. We used simulated gastric fluid (SGF) to simulate the human gastric environment. Three markers (rbcL, trnH-psbA, and ITS) were amplified and sequenced for all untreated and 15 mock-digested samples. The validity of DNA barcoding for species identification was assessed using the Basic Local Alignment Search Tool (BLAST) method and the tree construction method. The levels of three toxic components (saponin, phytoagglutin and trasylol) were determined in all samples using ELISA kits. The amplification success rate of all three regions was high (rbcL 96.67%, trnH-psbA 100%, and ITS 100%), but the sequencing of the trnH-psbA region was less satisfactory (66.67%), and SGF had a significant impact on the sequencing of the ITS region (After 40 min of SGF treatment, the sequencing success rate decreased by 46.67%). The samples from different species and origins contained different levels of toxic components, and the levels of all three substances decreased significantly after undergoing SGF digestion. After 1 h of SGF treatment, the saponin content decreased to 0–8.60% in untreated content (PHA decreased to 8.62–36.88%, trasylol decreased to 4.70–47.06%). The current results suggest that DNA barcoding has great potential for rapid identification of Leguminous poisoning in clinical settings. Toxins are probably not detectable in the patient for longer periods of poisoning. We recommend DNA barcoding technology as a first step for rapid screening and combined with toxin analysis for clinical diagnosis.
Similar content being viewed by others
Data availability
The dataset provided in this study can be found in online repositories. The name and accession number(s) of the repository can be found in the article or supplementary materials.
References
Azizi MMF, Lau HY, Abu-Bakar N (2021) Integration of advanced technologies for plant variety and cultivar identification. J Biosci.https://doi.org/10.1007/s12038-021-00214-x
Cahyaningsih R, Compton LJ, Rahayu S, Magos Brehm J, Maxted N (2022) DNA barcoding medicinal plant species from Indonesia. Plants (basel) 11(10):1375
Cavada BS, Osterne VJS, Oliveira MV, Pinto-Junior VR, Silva MTL, Bari AU, Lima LD, Lossio CF, Nascimento KS (2020) Reviewing mimosoideae lectins: a group of under explored legume lectins. Int J Biol Macromol 154:159–165
De Coninck T, Van Damme EJM (2021) Review: the multiple roles of plant lectins. Plant Sci 313:111096
Gostel MR, Zúñiga JD, Kress WJ, Funk VA, Puente-Lelievre C (2020) Microfluidic enrichment barcoding (MEBarcoding): a new method for high throughput plant DNA barcoding. Sci Rep 10(1):8701
Gupta RK, Gupta K, Sharma A, Das M, Ansari IA, Dwivedi PD (2017) Health risks and benefits of chickpea (Cicer arietinum) consumption. J Agric Food Chem 65(1):6–22
He S, Simpson BK, Sun H, Ngadi MO, Ma Y, Huang T (2018) Phaseolus vulgaris lectins: a systematic review of characteristics and health implications. Crit Rev Food Sci Nutr 58(1):70–83
Hebert PD, Cywinska A, Ball SL, deWaard JR (2003) Biological identifications through DNA barcodes. Proc Biol Sci 270(1512):313–321
Hui WF, Hon KL, Leung AKC (2021) An overview of the pediatric toxidromes and poisoning management. Curr Rev Clin Exp Pharmacol 16(4):318–329
Hui L, Jinghuan R, Yating W, Xiaoye W, Rui W (2022) Analysis of the epidemic characteristics of food poisoning incidents in China in 2018. Chin J Food Hyg 34(01):147–153
Jiao J, Jia X, Liu P, Zhang Q, Liu F, Ma C, Xi P, Liang Z (2018) Species identification of polygonati rhizoma in China by both morphological and molecular marker methods. C R Biol 341(2):102–110
Kaiqun L, Ling W, Yue F, Hui Y (2020) A case of toxic optic neuropathy in both eyes due to raw cowpea kernels. Chin J Ophthalmol Med 10(06):363–367
Karthikeyan A, Amalnath SD (2017) Abrus precatorius poisoning: a retrospective study of 112 patients. Indian J Crit Care Med 21(4):224–225
Kong X, Li Y, Liu X (2022) A review of thermosensitive antinutritional factors in plant-based foods. J Food Biochem 46(9):e14199
Krenzelok EP, Mrvos R (2011) Friends and foes in the plant world: a profile of plant ingestions and fatalities. Clin Toxicol (phila) 49(3):142–149
Lagarda-Diaz I, Guzman-Partida AM, Vazquez-Moreno L (2017) Legume lectins: proteins with diverse applications. Int J Mol Sci 18(6):1242
Lee EJ, Kim SC, Hwang IK, Yang HJ, Kim YS, Han MS, Yang MS, Lee YH (2009) The identification of ingested dandelion juice in gastric contents of a deceased person by direct sequencing and GC-MS methods. J Forensic Sci 54(3):721–727
Loera-Sánchez M, Studer B, Kölliker R (2020) DNA barcode trnH-psbA is a promising candidate for efficient identification of forage legumes and grasses. BMC Res Notes 13(1):35
Lonati D, Schicchi A, Crevani M, Buscaglia E, Scaravaggi G, Maida F, Cirronis M, Petrolini VM, Locatelli CA (2020) Foodborne botulism: clinical diagnosis and medical treatment. Toxins (basel) 12(8):509
Mudryj AN, Yu N, Aukema HM (2014) Nutritional and health benefits of pulses. Appl Physiol Nutr Metab 39(11):1197–1204
Naithani S, Komath SS, Nonomura A, Govindjee G (2021) Plant lectins and their many roles: carbohydrate-binding and beyond. J Plant Physiol 266:153531
Ng AE, Sandoval E, Murphy TM (2016) Identification and individualization of Lophophora using DNA analysis of the trnL/trnF region and rbcL gene. J Forens Sci 61(Suppl 1):S226-229
Ninan EC, James E (2019) Acute disseminated encephalomyelitis due to abrus precatorius poisoning—a case report. Saudi Pharm J 27(4):521–524
Robinson GHJ, Domoney C (2021) Perspectives on the genetic improvement of health- and nutrition-related traits in pea. Plant Physiol Biochem 158:353–362
Romano MC, Doan HK, Poppenga RH, Filigenzi MS, Bryant UK, Gaskill CL (2019) Fatal amanita muscaria poisoning in a dog confirmed by PCR identification of mushrooms. J Vet Diagn Invest 31(3):485–487
Singh B, Singh JP, Singh N, Kaur A (2017) Saponins in pulses and their health promoting activities: a review. Food Chem 233:540–549
Tor-Roca A, Garcia-Aloy M, Mattivi F, Llorach R, Andres-Lacueva C, Urpi-Sarda M (2020) Phytochemicals in Legumes: a qualitative reviewed analysis. J Agric Food Chem 68(47):13486–13496
Wei S, Luo Z, Cui S, Qiao J, Zhang Z, Zhang L, Fu J, Ma X (2019) Molecular identification and targeted quantitative analysis of medicinal materials from uncaria species by DNA barcoding and LC-MS/MS. Molecules 24(1):175
Xie L, Wang YW, Guan SY, Xie LJ, Long X, Sun CY (2014) Prospects and problems for identification of poisonous plants in China using DNA barcodes. Biomed Environ Sci: BES 27(10):794–806
Yun L, Zhipeng M, Yang Z, Min D, Chaoxia G, Hao P, Xueqin S, Dengmei X (2022) Epidemic characteristics and disposal of bean food poisoning events in Kunming from 2004 to 2019. Med Inform 35(03):143–145
Zhang R, Qin X, Zhang T, Li Q, Zhang J, Zhao J (2018) Astragalus polysaccharide improves insulin sensitivity via AMPK activation in 3T3-L1 adipocytes. Molecules 23(10):2711
Zhang L, Lv Q, Zheng Y, Gao S, Huang W, Liu P, Kong D, Wang Y, Yu Y, Jiang Y, Jiang H (2022) Rapid and sensitive detection of botulinum toxin type A in complex sample matrices by AlphaLISA. Front Public Health 10:987517
Zhu S, Liu Q, Qiu S, Dai J, Gao X (2022) DNA barcoding: an efficient technology to authenticate plant species of traditional Chinese medicine and recent advances. Chin Med 17(1):112
Acknowledgements
The financial support of Qingdao Municipal Center for Disease Control and Prevention for the preparation of this manuscript is greatly appreciated.
Funding
This project was supported by the Qingdao Minsheng Science and Technology Plan Project (18–6-1–67-nsh).
Author information
Authors and Affiliations
Contributions
Jie Wang conceived and designed the study. Xiaojing Lu, Hongwei Yu, Jinquan Zhao, Shuhua Bu, and Ze Liu performed the experiments. Wisen Yu provided funding and experimental equipment. Jie Wang and Shuangyu Wang wrote the manuscript. Fenglin Sun and Chang Liu compiled the experimental data and revised the manuscript. All the authors read and approved the final manuscript.
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Wang, J., Wang, S., Sun, F. et al. Molecular identification of DNA barcoding of Leguminous toxic species and quantitative analysis by ELISA kits. Plant Biotechnol Rep 18, 233–241 (2024). https://doi.org/10.1007/s11816-024-00892-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11816-024-00892-7